Literature DB >> 29379330

Biological activities of the red algae Galaxaura rugosa and Liagora hawaiiana butters.

Nouf M Al-Enazi1, Amani S Awaad2, Saleh I Alqasoumi3, Metab F Alwethairi4.   

Abstract

The biological activities; antimicrobial, antioxidant and anticancer, of the red algae Galaxaura rugosa and Liagora hawaiiana were determined. The total ethanol, lipoidal matters, chloroform, n-butanol, aqueous extracts and powder of both algae showed and bacterial and antifungal activities. However, the chloroform extract of Galaxaura rugosa showed antibacterial activity against Klebsiella pneumoniae (24 mm, 0.15 mg/ml) higher than gentamycin (23 mm, 0.49 mg/ml). Moreover, the total ethanol, lipoidal matter and chloroform extracts showed antifungal activity (21, 22 and 25 mm, 1.25, 0.312 and 0.156 mg/ml) similar to the antibiotic Ketoconazole activity (23, 24 and 27 mm, 1.25, 0.312 and 0.156 mg/ml) against Aspergillus fumigatus, A. niger and Candida trobicalis, respectively. A good antioxidant activity (80.96%, IC50 = 27.8 µg/ml) was provided by Galaxaura rugosa. The anticancer activity results revealed that the lipoidal matters of Galaxaura rugosa and Liagora hawaiiana possessed antitumor activity (IC50 = 15 ± 1.7 and 21.2 ± 1.6, respectively) against lung carcinoma (A-549) better than vinblastine sulfate (IC50 = 24.6 ± 0.7). Although, the lipoidal matters of Galaxaura rugosa and Liagora hawaiiana antitumor activity against cervical carcinoma (HeLa) and intestinal carcinoma (CACO-2) (IC50 = 10.2 ± 0.6 and 12.2 ± 0.6, respectively) preferable than vinblastine sulfate (IC50 = 59.7 ± 2.1 and 30.3 ± 1.4, respectively).

Entities:  

Keywords:  Antimicrobial; Antioxidant; Antitumor; Extraction; Phytochemical screening

Year:  2017        PMID： 29379330      PMCID： PMC5783822          DOI： 10.1016/j.jsps.2017.11.003

Source DB:  PubMed          Journal:  Saudi Pharm J        ISSN： 1319-0164            Impact factor:   4.330


Introduction

The interest in ancient herbal remedies has been significantly increased in the last few decades. In the worldwide, all the natural resources including medicinal plants, fungi and algae are screened for their biological activities (Awaad et al., 2013, Zain et al., 2012, Amornlerdpison et al., 2007). Accordingly, the therapeutic values and pharmaceutical usage of numerous herbal medicines have already been validated. The herbal medicines which obtained from natural sources are considered as safe for human beings. However, they would have some antagonistic effects due to presence of other active ingredients (Izzo and Ernst, 2009). Algae are found everywhere: in the sea, rivers, lakes, soil, walls, and as symbiont in animal and plants. Algae include four main divisions; namely, Red algae (Rhodophya), Brown Algae (Phycophyta), Green Algae (Chlorophyta) and Diatoms. Although, Seaweeds which are macroscopic, multicellular, and marine algae, are divided into three categories; red, green and brown organisms comprises about 30000 species. In most of Asian countries, seaweeds are traditionally traded as food items including sushi wrappings, seasonings, condiments, and vegetables (El El Gamal, 2010, Mark et al., 2016). Antioxidants have attracted the most interest among the many biologically-active compounds found in algae. Antioxidants are important compounds in the treatment and recovery from various diseases including cancer, chronic inflammation, atherosclerosis, cardiovascular disorders, and aging process (Kohen and Nyska, 2002). Although, the search for anticancer drugs has similar attention as marine compounds revealed promising results at different stages of cancer progress (Mayer and Gustafson, 2006). On the other hand, in developed and developing countries, the most people died following infectious bacterial and/or fungal diseases. The bacterial Gram-positive and Gram-negative organisms including different species of Bacillus, Proteus, Klebsiella, Staphylococcus, Salmonella and Pseudomonas are the main source of severe infections in animals including humans (Nathan, 2004). Among seaweeds, numerous macroalgae have potent cytotoxic activities (Mayer and Gustafson, 2006, Smit, 2004) and algal consumption has been suggested as a chemo-preventive agent against several cancers (Yuan and Walsh, 2006). Recently, due to their exceptional richness in bioactive compounds (e.g., antimicrobial, anti-inflammatory, and antitumoral activities), the seaweeds has significantly expanded into the pharmaceutical and para-pharmaceutical industry (Kornprobst, 2005, Smit, 2004). The current study aimed to assess the biological activity including antioxidant, antimicrobial, and anticancer of different extracts of the red algae Galaxaura rugosa and Liagora hawaiiana.

Material and methods

Algal samples collection, extraction and screening

Algal species collections

The algal species used in this study; namely, Galaxaura rugose and Liagora hawaiiana Butters were collected from Alharra, Umluj, Red Seashore, Kingdom of Saudi Arabia. Algal species were identified according to Aleem, 1993, Coppejans et al., 2009. Samples collected were air-dried in shade, reduced to fine powder, packed in tightly closed containers and stored for phytochemical and biological studies.

Algal extraction

Dry powder (830 and 795 g) of Galaxaura rugose and Liagora hawaiiana; respectively, were extracted by percolation in 95% ethanol (Awaad et al., 2017a) at room temperature for two days. The total ethanol extract was filtered and the residue was re-percolated by the same manor for five times. The ethanol extract was then concentrated, under reduced pressure at low temperature, and a yield of 81 and 77 g was obtained from Galaxaura rugose and Liagora hawaiiana, respectively. The obtained extracts of each algae was separately suspended in water (300 ml) and filtered over a piece of cotton. The lipoidal matter, collected on top of the cotton piece (25 and 28 g. for Galaxaura rugose and Liagora hawaiiana, respectively) were obtained. The aqueous layer, which filtered off, was successively fractionated using chloroform and n-butanol. Each extract was dried over anhydrous sodium sulfate, concentrated and yielded 11 & 30 g and 14 and 26 g for chloroform and n-butanol of Galaxaura rugos and Liagora hawaiiana, respectively. However, after extraction with n-butanol some powder was precipitated from each algae and the filtration was carried out to separate it and. The leftover aqueous extract of each alga was dried using lyophilization (Awaad et al., 2017b) and kept for further investigation.

Phytochemical screening

Powdered sample of each investigated alga (Galaxaura rugose and Liagora hawaiiana) was subjected to phytochemical screening as published by Khan et al. (2011) to investigate their phytochemical constituents.

Antimicrobial activity

Test organisms

Different clinically isolated bacterial and fungal strains; namely, Aspergillus fumigatus (RCMB 02568), Aspergillus niger (RCMB 02724), Bacillus substilis (RCMB 010015), Candida albicans (RCMB 05003), Candida. tropicalis (RCMB 05004), Cryptococcus neoformans (RCMB 05642), Escherichia coli (RCMB 010052), Geotricum candidum (RCMB 05097), Klebsiella pneumonia (RCMB 0010093), Microsporum canis (RCMB 0834), Penicillium expansum (RCMB 01924), Pseudomonas aeruginosa (RCMB 0100243-5), Proteous vlgaris (RCMB 01004) Staphylococcus aureus (RCMB 010010), Staphylococcus epidermidis (RCMB 010009), Streptococcus byogenes (RCMB 0100174-2), Stroptococcus mutans (RCMB 0100017) Salmonella typhimurium, RCMB (RCMB 14028), Syncephalastrum racemosum (RCMB 05922) and Trichophyton mentagrophytes (RCMB 0925) were obtained from the Microbiology Laboratory, Regional Center for Mycology and Biotechnology, Al-Azhar University, Cairo, Egypt and used as test organisms.

Antimicrobial assay

The antibacterial and antifungal activities of total ethanol, lipoidal matters, chloroform n-butanol, aqueous extracts and powder of Galaxaura rugosa and Liagora hawaiiana were determined using the well-diffusion method (Almalki, 2017). Petri plates containing 20 ml of, nutrient (for bacteria) or malt extract (for fungi), agar medium were seeded with 1–3 day cultures of microbial inoculums. Wells (6 mm in diameter) were cut off from agar and 50 µl of algal extracts were tested in a concentration of 100 mg/ml and incubated at 37 °C for 24–48 h (bacterial strains) and for 3–5 days (fungal strains). The antibacterial and antifungal activities were determined by measurement of the diameter of the inhibition zone around the well.

Determination of minimum inhibitory concentration (MIC)

The minimum inhibitory concentration (MIC) of algal extract was determined by micro-dilution method using serially diluted (2 folds) algal extracts (Zain et al., 2012). The MIC of total ethanol, lipoidal matter, chloroform, n-butanol, aqueous extracts and powder of Galaxaura rugosa and Liagora hawaiiana were determined by dilution of concentrations from 0.0 to 100 mg/ml. Equal volumes of each extract and nutrient broth were mixed in a test tube. Specifically 0.1 ml of standardized inoculum (1–2 × 107 cfu/ml) was added in each tube. The tubes were incubated at 37 °C for 24–48 h and/or 3–5 days. Two control tubes, containing the growth medium, saline and the inoculum were maintained for each test batch. The lowest concentration (highest dilution) of the algal extract that produced no visible microbial growth (no turbidity) when compared with the control tubes were regarded as MIC.

Antioxidant assay

The antioxidant activity of Galaxaura rugosa and Liagora hawaiiana different extracts were determined using DPPH free radical scavenging assay as describe by Aksoy et al. (2013) in triplicate and average values were considered. The tested extracts were also compared using the IC50 value; i.e., the concentration leading to 50% inhibition which was estimated from graphical plots of DPPH Radical Scavenging% Vs concentrations.

Antitumor activity

The antitumor activity of total ethanol, lipoidal matters, chloroform, n-butanol, aqueous extracts and powder of Galaxaura rugosa and Liagora hawaiiana were determined using A-549 (Lung carcinoma), CACO (colorectal carcinoma), HCT-116 (Colon carcinoma), Hela (Cervical carcinoma), HEp-2 (Larynx carcinoma), HepG-2 (Hepatocellular carcinoma), and MCF-7 (Breast carcinoma) cell lines as described by Kameyama et al. (2005).

Statistical analysis

All values were expressed as mean ± S.D. Comparisons between means were carried out using a one-way ANOVA test followed by the Tukey HSD test using SPSS, version 14 (SPSS, Chicago, IL). Differences at p50.05 were considered statistically significant.

Results and discussion

Preliminary phytochemical screening

The preliminary phytochemical analyses of Galaxaura rugosa and Liagora hawaiiana revealed the presence of different primary and secondary metabolites, they contains unsaturated sterols and/or triterpenoids, flavonoids, carbohydrates or glycosides, proteins and/or amino acids, tannins and coumarin, no saponins or alkaloids were detected. This variety of active metabolites give these algae high potentials to be used as source of medication specially the presence of flavonoids (Kosanić et al., 2015). The antimicrobial activity of total ethanol, lipoidal matters, chloroform, n-butanol, aqueous extracts, and powder of Galaxaura rugosa and Liagora hawaiiana were determined against Gram-negative, Gram-positive bacteria and fungi (Table 1, Table 2). The results revealed that all the extracts of Galaxaura rugosa showed antibacterial and antifungal activities. On the other hand, only lipoidal matters, chloroform, n-butanol and aqueous extracts of Liagora hawaiiana showed antibacterial and antifungal activity, in addition to the powder which has only antifungal activity (Table 1, Table 2).
Table 1

Antimicrobial activity of different extracts of Galaxaura rugosa.

ExtractTest organismMean diameter of inhibition zone (mm)
Total (Ethanol)Lipoidal matterChloroformn-ButanolAqueousPowderStandard antibiotic
BacteriaGram-negativeGentamycin
Escherichia coli15182216221536
Klebsiella pneumoniae14192415161423
Proteous vulgaris18222019151631
Pseudomonas aeruginosa00001500000025
Salmonella typhimrium00142115150027



Gram-positive
Bacillus substilis14181716161532
Staphylococcus aureus19142221182030
Staphylococcus epidermidis20001921171434
Streptococcus mutans00001415200026
Streptococcus pyogenes00000014180028



FungiKetocona-zole
Aspergillus fumigatus21160000000023
Aspergillus niger15220000000024
Candida albicans18162018151826
Candida trobicalis17192519162327
Cryptococcus neoformans20152722181531
Geotricum candidum15172220152230
Penicillium expansum14230000000028
Syncephalastrum racemosum14140000000024



DermatophytesAmphotericin B
Microsporum canis00000000000030
Trichophyton mentagrophytes00000000000029

Values are expressed as mean ± SEM of 3 determinants.

Table 2

Antimicrobial activity of different extracts of Liagora hawaiiana.

ExtractTest organismMean diameter of inhibition zone (mm)
Total(Ethanol)Lipoidal matterChloroformn-ButanolAqueousPowderStandard Antibiotic
BacteriaGram-negativeGentamycin
Escherichia coli00212115230036
Klebsiella pneumoniae00222016160023
Proteous vulgaris00151821150031
Pseudomonas aeruginosa00141500000025
Salmonella typhimrium00191614000027
Gram-positive
Bacillus substilis00151700000032
Staphylococcus aureus00171800000030
Staphylococcus epidermidis00141400000034
Streptococcus mutans00202400000026
Streptococcus pyogenes00191900000028



FungiKetocona-zole
Aspergillus fumigatus00001600000023
Aspergillus niger00001600000024
Candida albicans00212214161426
Candida trobicalis00222715181527
Cryptococcus neoformans00242519211731
Geotricum candidum00212114151430
Penicillium expansum00002000000028
Syncephalastrum racemosum00001500000024



DermatophytesAmphotericin B
Microsporum canis00000000000030
Trichophyton mentagrophytes00000000000029

Values are expressed as mean ± SEM of 3 determinants

Antimicrobial activity of different extracts of Galaxaura rugosa. Values are expressed as mean ± SEM of 3 determinants. Antimicrobial activity of different extracts of Liagora hawaiiana. Values are expressed as mean ± SEM of 3 determinants Among the extracts of Galaxaura rugosa, chloroform, n-butanol, and aqueous extracts inhibited the growth of nine, out of ten, bacterial test organism. While total ethanol extract and lipoidal matters showed antifungal activity against 8, out of ten, fungal test strains. Interestingly, the chloroform extract of Galaxaura rugosa exhibited antibacterial activity against Klebsiella pneumoniae (24 mm, 0.15 mg/ml) higher than the standard antibiotic Gentamycin (23 mm, 0.49 mg/ml). Moreover, the total ethanol, lipoidal matter and chloroform extracts showed antifungal activity (21, 22 and 25 mm, 1.25, 0.312 and 0.156 mg/ml) similar to the antibiotic Ketoconazole activity (23, 24 and 27 mm, 1.25, 0.312 and 0.156 mg/ml) against Aspergillus fumigatus, A. niger and Candida trobicalis, respectively (Table 1, Table 3). The chloroform extract of Liagora hawaiiana showed the best antibacterial and antifungal activities. With the exception of Microcanis canis and Trichophyton mentagrophytes, it inhibited the growth of all tested fungal strains in addition to all the bacterial strains. Furthermore, the potency of chloroform extract against Candida tropicalis (27 mm, 0.078 mg/ml) was similar to that of the standard antibiotic, Ketoconazole (27 mm, 0.98 mg/ml) (Table 2, Table 4).
Table 3

The minimum inhibitory concentration (MIC) of different extracts of Galaxaura rugosa.

ExtractTest organismMinimum Inhibitory Concentration (mg/ml)
Total (Ethanol)Lipoidal matterChloroformn-ButanolAqueousPowderStandard Antibiotic
BacteriaGram-negativeGentamycin
Escherichia coli5.0002.5000.3125.0000.6255.00003.90
Klebsiella pneumoniae10.002.5000.1565.0005.00010.0000.49
Proteous vulgaris2.5000.6251.2502.5005.0005.00001.95
Pseudomonas aeruginosaNDND5.000NDNDND01.95
Salmonella typhimriumND10.000.6255.0005.000ND01.95



Gram-positive
Bacillus substilis10.001.2502.5005.0005.0005.00001.95
Staphylococcus aureusND10.000.6250.6252.5001.25001.95
Staphylococcus epidermidisNDND1.2500.6252.50010.00000.98
Streptococcus mutansNDND10.005.0001.250ND01.95
Streptococcus pyogenesNDNDND10.002.500ND00.98



FungiKetocona-zole
Aspergillus fumigatus1.2505.000NDNDNDND00.49
Aspergillus niger5.0000.312NDNDNDND03.90
Candida albicans2.5005.0001.2501.2505.0001.25001.95
Candida trobicalis2.5001.2500.1562.5005.0000.31200.98
Cryptococcus neoformans1.2505.0000.0780.6252.5005.00001.95
Geotricum candidum5.0002.5000.3121.2505.0000.62503.90
Penicillium expansum10.000.312NDNDNDND01.95
Syncephalastrum racemosum10.0010.00NDNDNDND00.98

ND, not determined. Values are expressed as mean ± SEM of 3 determinants.

Table 4

The minimum inhibitory concentration (MIC) of different extracts of Liagora hawaiiana.

Extract TestorganismMinimum Inhibitory Concentration (mg/ml)
Total (Ethanol)Lipoidal matterChloroformn-ButanolAqueousPowderStandard antibiotic
BacteriaGram-negativeGentamycin
Escherichia coliND0.6250.6255.0000.625ND03.90
Klebsiella pneumoniaeND0.3121.2505.0005.000ND00.49
Proteous vulgarisND10.002.5000.62510.00ND01.95
Pseudomonas aeruginosaND10.005.000NDNDND01.95
Salmonella typhimriumND1.2505.00010.00NDND01.95
Gram-positive
Bacillus substilisND5.0002.500NDNDND01.95
Staphylococcus aureusND2.5001.250NDNDND01.95
Staphylococcus epidermidisND10.0010.00NDNDND00.98
Streptococcus mutansND1.2500.312NDNDND01.95
Streptococcus pyogenesND2.5001.250NDNDND00.98



FungiKetocona-zole
Aspergillus fumigatusNDND5.000NDNDND00.49
Aspergillus nigerNDND10.00NDNDND03.90
Candida albicansND0.6250.62510.005.00010.0001.95
Candida trobicalisND0.3120.0785.0002.5005.00000.98
Cryptococcus neoformansND0.1560.3121.2500.6255.00001.95
Geotricum candidumND0.6250.62510.005.00010.0003.90
Penicillium expansumNDND1.250NDNDND01.95
Syncephalastrum racemosumNDND5.000NDNDND00.98

ND, not determined. Values are expressed as mean ± SEM of 3 determinants.

The minimum inhibitory concentration (MIC) of different extracts of Galaxaura rugosa. ND, not determined. Values are expressed as mean ± SEM of 3 determinants. The minimum inhibitory concentration (MIC) of different extracts of Liagora hawaiiana. ND, not determined. Values are expressed as mean ± SEM of 3 determinants. From the previous studied it was concluded that researchers have isolated different compounds from algae including terpenoids, phlorotannins, polyphenols, phenolic acids, anthocyanins, hydroxycinnamic acid derivatives, and flavonoids (Bhat and Madyastha, 2000, Bhat and Madyastha, 2001, Benedetti et al., 2004). Nevertheless, the antibacterial, antifungal and antiviral activities of algal extracts are extensively published (El-Fatemy and Said, 2011, Manilal et al., 2009, Rajasulochana et al., 2009, Ely et al., 2004). Although, the obtained results of the current study revealed the antimicrobial activity of extracts of Galaxaura rugosa and Liagora hawaiiana using different solvents which indicates the multiplicity and diversity of the compounds present in algae

Antioxidant activity

The antioxidant activity of Galaxaura rugosa, and Liagora hawaiiana were screened using DPPH assay. It is the most commonly used assay because it can run many samples in short time and detect the active components at low concentration (Piao et al., 2004). The current results exhibited that the total ethanol extract of Galaxaura rugosa, and Liagora hawaiiana have DPPH radical scavenging activity in a concentration–dependent manner (Table 5). The maximum scavenging activity (80.96%, IC50 = 27.8 µg/ml) was provided by Galaxaura rugosa. However, the scavenging activity of Liagora hawaiiana was 66.87% (IC50 = 57.2 µg/ml) (Table 5).
Table 5

The scavenging activity of DPPH radicals of Galaxaura rugosa and Liagora hawaiiana.

Concentration (µg/ml)DPPH scavenging (%)
Galaxaura rugosaLiagora hawaiianaAscorbic acid
00000.0000.0000.00
00110.87 ± 1.504.96 ± 1.3212.98 ± 1.41
00212.35 ± 1.119.83 ± 1.2116.38 ± 1.44
00421.39 ± 1.7116.61 ± 1.5462.98 ± 1.62
00828.09 ± 1.3222.35 ± 1.3376.81 ± 1.57
01634.35 ± 1.9127.91 ± 1.3878.72 ± 1.75
03255.48 ± 1.2235.65 ± 1.3078.94 ± 1.51
06466.00 ± 1.5853.83 ± 1.2780.21 ± 1.14
12880.96 ± 1.3066.87 ± 1.1286.36 ± 1.09
IC5027.8 ± 1.2257.2 ± 1.3511.2 ± 1.55

Values are expressed as mean ± SEM of 3 replicates.

The scavenging activity of DPPH radicals of Galaxaura rugosa and Liagora hawaiiana. Values are expressed as mean ± SEM of 3 replicates. The free radicals are involved in several diseases including cancer, AIDS and neurodegenerative diseases. The scavenging activity of antioxidants is very useful for the control of those diseases (Suresh et al., 2008, Kohen and Nyska, 2002). Interestingly, the antioxidant activity of Galaxaura rugosa was very good (27.8 ± 1.22) and almost similar to the antioxidant activity of ascorbic acid (86.36%, IC50 = 11.2 µg/ml) (Table 5), this can be due to the presence of flavonoids in both algae (Farasat et al., 2014, Yen and Duh, 1994). The cancer, cells growing out of control, causes are diverse, complex and not fully understood. The cancer diseases are classified according to the type of cell that the tumor cells resemble and are presumed to be the origin of the tumor. Herbal medicines are used worldwide for cancer prevention and treatment. The effect of natural products as anti-cancer was widely studied because their nature, low toxicity and side effects (Manglani et al., 2014, Mulla and Swamy, 2012, Jain and Jain, 2011). In the present study, the in vitro antitumor activity of Galaxaura rugosa and Liagora hawaiiana extracts was determined against different cell lines including A-549 (Lung carcinoma), CACO (Intestinal carcinoma), HCT-116 (Colon carcinoma), Hela (Cervical carcinoma), HEp-2 (Larynx carcinoma), HepG-2 (Hepatocellular carcinoma), and MCF-7 (Breast carcinoma). Because it is reliable to assess the in vitro cytotoxicity of the anticancer compounds, MTT assay method (Allely et al., 1998) was used. The obtained results revealed that the extracts of Galaxaura rugosa and Liagora hawaiiana have a remarkable antitumor activity against different types of tumor cells (Table 6, Table 7). Interestingly, the lipoidal matters of Galaxaura rugosa and Liagora hawaiiana possessed antitumor activity (IC50 = 15±1.7 and 21.2 ± 1.6, respectively) against lung carcinoma (A-549) better than vinblastine sulfate (IC50 = 24.6 ± 0.7). Although, the lipoidal matters of Galaxaura rugosa and Liagora hawaiiana antitumor activity against cervical carcinoma (HeLa) and intestinal carcinoma (CACO-2) (IC50 = 10.2 ± 0.6 and 12.2 ± 0.6, respectively) preferable than vinblastine sulfate (IC50 = 59.7 ± 2.1 and 30.3 ± 1.4, respectively) (Table 6, Table 7). These results give new promising resource of anticancer drug discovery from marine this was clear from the variation of the anticancer effect of the algae extracts which due to their huge biodiversity and safety, as they have long been used in traditional Asian foods and folk medicine (Namvar et al., 2014)
Table 6

The antitumor activity of extracts of Galaxaura rugosa against different cell lines.

Cell lineConcentration (µg/ml)Cell viability (%)
Total (Ethanol)Lipoidal mattersChloro-formn-butanolAqueousPowderVinblastine sulfate
A-549Lung carcinoma000.00100100100100100100100
001.0010098.110010010010098.2
002.0098.192.310010010010094.7
003.9094.084.998.610010098.481.4
007.8087.367.293.710098.191.773.8
015.6076.948.685.199.292.885.062.5
031.2563.140.970.495.084.072.340.7
062.5041.932.854.889.469.559.132.9
125.0030.621.938.768.142.738.625.2
250.0022.812.823.840.729.423.115.3
500.0010.706.312.927.814.510.906.8



CACO-2Intestinal carcinomaIC50 (µg/ml)50.7 ± 3.515 ± 1.781.4 ± 4.5208 ± 17.2108 ± 9.290.4 ± 7.824.6 ± 0.7
000.00100100100100100100100
001.0010010010010010010099.2
002.0010098.710010010010093.8
003.9010095.410010097.810086.2
007.8099.489.299.497.492.499.479.4
015.6096.172.395.290.685.296.267.5
031.2589.249.284.181.478.189.748.9
062.5072.538.470.668.065.770.831.4
125.0043.827.153.447.142.942.920.3
250.0031.714.234.936.228.728.808.9
500.0016.406.423.621.412.515.204.0
IC50 (µg/ml)112 ± 10.430.7 ± 4.1149 ± 12.2117 ± 9.1106 ± 8.4109 ± 11.430.3 ± 1.4



HCT-116Colon carcinoma000.00100100100100100100100
001.0010079.410010010010066.4
002.0010072.910010010010058.1
003.9098.760.710010010098.747.3
007.8093.245.999.198.110095.139.8
015.6086.938.293.791.899.789.528.7
031.2569.130.686.085.292.473.118.9
062.5043.522.868.168.069.549.515.5
125.0030.716.445.249.842.836.912.1
250.0018.608.731.735.424.923.806.7
500.0006.03.918.623.808.709.204.0
IC50 (µg/ml)54.7 ± 1.26.7 ± 0.2112 ± 7.2125 ± 5.3108 ± 3.961.9 ± 3.13.5 ± 0.2



HeLaCervical carcinoma000.00100100100100100100100
001.0010092.5100100100100100
002.0099.581.410010010010098.1
003.9094.669.099.410010098.495.4
007.8089.752.796.010097.892.390.6
015.6080.943.988.910091.381.482.7
031.2565.235.180.710080.669.871.9
062.5048.126.768.910062.945.147.8
125.0031.418.447.510043.030.634.5
250.0014.709.631.797.129.421.822.8
500.0008.905.717.289.215.809.209.1
IC50 (µg/ml)59.1 ± 3.210.2 ± 0.6118 ± 5.1> 500103 ± 4.856.3 ± 3.459.7 ± 2.1



HepG-2Hepatocellular carcinoma000.00100100100100100100100
001.0010090.610010010010060.9
002.0098.283.110010010098.754.2
003.9091.770.499.310098.694.045.0
007.8080.149.294.110091.788.734.1
015.6069.437.086.210082.072.126.8
031.2548.128.572.610067.448.519.2
062.5034.520.651.310053.936.414.3
125.0023.711.334.999.037.124.910.9
250.0011.906.926.493.721.315.305.8
500.0005.603.513.881.409.406.103.2



MCF-7Breast carcinomaIC50 (µg/ml)29.9 ± 2.37.6 ± 0.567.6 ± 4.2> 50077.2 ± 5.930.3 ± 2.62.9 ± 0.3
000.00100100100100100100100
001.0010094.110010010010067.1
002.0010089.210010010010058.7
003.9099.478.310010010099.452.9
007.8096.263.110010010096.247.2
015.6089.442.598.110098.789.540.5
031.2572.331.789.710092.470.831.9
062.5040.923.874.010078.141.723.8
125.0026.414.748.798.745.228.515.1
250.0013.807.532.891.430.918.707.8
500.0006.703.819.476.813.708.905.4
IC50 (µg/ml)53.5 ± 2.312.8 ± 1.4122 ± 9.3>500116 ± 8.253.6 ± 4.65.9 ± 0.4



PC-3Prostate carcinoma000.00100100100100100100100
001.0010010010010010010093.0
002.0010010010010010010088.2
003.9098.010010010010010074.8
007.8091.797.810010098.710068.9
015.6086.990.610010093.810056.7
031.2571.479.597.610084.110037.8
062.5043.564.093.899.567.299.424.9
125.0029.438.185.192.443.996.213.7
250.0017.227.869.483.928.688.409.5
500.0008.215.846.270.215.874.505.3
IC50 (µg/ml)55.3 ± 5.496.4 ± 8.3459 ± 24.8> 500109 ± 7.8> 50021.2 ± 0.9

Values are expressed as mean ± SEM of 3 determinants.

Table 7

The antitumor activity of extracts of Liagora hawaiiana against different cell lines.

Cell lineConcentration (µg/ml)Cell viability (%)
Total (Ethanol)Lipoidal mattersChloro-formn-butanolAqueousPowderVinblastine sulfate
A-549Lung carcinoma000.00100100100100100100100
001.0010099.698.710010010098.2
002.0099.894.895.210010010094.7
003.9096.789.586.710010010081.4
007.8090.370.479.510098.110073.8
015.6080.656.263.898.091.498.762.5
031.2568.538.740.886.979.594.340.7
062.5053.228.131.972.163.887.132.9
125.0034.922.423.550.645.969.225.2
250.0021.814.014.839.837.651.815.3
500.0009.7906.106.926.524.932.706.8



CACO-2Intestinal carcinomaIC50 (µg/ml)73.6 ± 5.821.2 ± 1.625 ± 3.1132 ± 11.4111 ± 8.9274 ± 26.224.6 ± 0.7
000.00100100100100100100100
001.0010093.710010010010099.2
002.0010089.499.498.110010093.8
003.9010078.196.391.799.710086.2
007.8098.065.387.286.493.810079.4
015.6090.638.470.975.385.298.367.5
031.2578.128.937.460.869.192.548.9
062.5065.920.428.645.148.781.431.4
125.0047.813.219.436.239.465.120.3
250.0031.706.710.523.627.841.808.9
500.0018.603.205.911.715.930.604.0
IC50 (µg/ml)118 ± 10.512.2 ± 0.625.4 ± 1.252.9 ± 3.360.5 ± 4.7206 ± 16.230.3 ± 1.4



HCT-116Colon carcinoma000.00100100100100100100100
001.0010096.791.410010010066.4
002.0098.190.686.210010010058.1
003.9094.283.978.110099.410047.3
007.8086.170.863.998.295.210039.8
015.6071.852.350.692.488.799.428.7
031.2559.442.941.086.979.495.218.9
062.5036.733.931.772.865.986.115.5
125.0021.820.622.445.141.870.912.1
250.0012.913.212.936.228.742.106.7
500.0005.606.305.220.416.228.604.0
IC50 (µg/ml)44.2 ± 0.9195 ± 0.716.6 ± 0.8114 ± 9.2104 ± 8.7216 ± 12.33.5 ± 0.2



HeLaCervical carcinoma000.00100100100100100100100
001.00100100100100100100100
002.0010099.798.610010010098.1
003.9097.896.493.210010010095.4
007.8090.390.685.410010010090.6
015.6078.481.776.498.697.910082.7
031.2563.169.462.191.790.610071.9
062.5048.251.842.583.178.210047.8
125.0031.536.730.470.865.199.734.5
250.0019.723.919.556.447.292.322.8
500.0008.314.508.738.631.581.609.1
IC50 (µg/ml)58.8 ± 1.470.2 ± 3.550.7 ± 2.9340 ± 16.7231 ± 20.1> 50059.7 ± 2.1



HepG-2Hepatocellular carcinoma000.00100100100100100100100
001.0010010099.810010010060.9
002.0010099.293.110010010054.2
003.9098.593.184.010098.710045.0
007.8090.185.769.510093.110034.1
015.6078.076.946.299.286.210026.8
031.2565.460.832.894.075.310019.2
062.5040.643.125.688.661.498.114.3
125.0028.330.414.373.143.091.810.9
250.0014.618.706.946.231.780.705.8
500.0006.810.203.428.919.468.903.2



MCF-7Breast carcinomaIC50 (µg/ml)50.8 ± 5.150.4 ± 4.314.4 ± 0.8233 ± 19.6101 ± 7.8> 5002.9 ± 0.3
000.00100100100100100100100
001.0010010097.210010010067.1
002.0010010091.710010010058.7
003.9010010085.010010010052.9
007.8099.597.076.910099.310047.2
015.6091.489.560.810095.110040.5
031.2579.871.339.699.489.510031.9
062.5045.149.828.196.570.898.223.8
125.0032.736.219.481.447.294.515.1
250.0019.421.408.763.135.983.107.8
500.0010.213.804.237.826.571.505.4
IC50 (µg/ml)58.1 ± 3.762.2 ± 6.123.6 ± 3.4380 ± 17.9118 ± 82.3> 5005.9 ± 0.4



PC-3Prostate carcinoma000.00100100100100100100100
001.0010010010010010010093.0
002.0010098.698.010010010088.2
003.9010091.791.710010010074.8
007.8010084.384.110098.710068.9
015.6098.068.170.810090.610056.7
031.2590.647.241.599.582.198.437.8
062.5072.835.023.793.167.491.324.9
125.0047.823.619.586.446.280.113.7
250.0031.714.910.271.635.965.309.5
500.0018.906.306.346.821.341.905.3
IC50 (µg/ml)120 ± 9.329.2 ± 1.326.7 ± 1.4469 ± 38.6114 ± 10.5414 ± 43.121.2 ± 0.9

Values are expressed as mean ± SEM of 3 determinants.

The antitumor activity of extracts of Galaxaura rugosa against different cell lines. Values are expressed as mean ± SEM of 3 determinants. The antitumor activity of extracts of Liagora hawaiiana against different cell lines. Values are expressed as mean ± SEM of 3 determinants.
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