Wa Zhong1, Fang-ju Kan, Zhong Yu. 1. Department of Digestive Diseases, The Second Affiliated Hospital of Sun Yat-sen University, Guangzhou.
Abstract
OBJECTIVE: To study the effect of Weikangning (WKN) containing serum on growth and cell cycle regulators of gastric cancer cell. METHODS: WKN containing drug serum was prepared by gastric perfusion of WKN in different dosages to SD rats. Gastric cancer cells were cultured in medium contained the drug serum with different concentrations of WKN. The change of cell cycle was detected by flow cytometry, and the mRNA and protein expressions of CyclinD2, CyclinE, Cyclin-dependant kinase2 (CdK2), Cdk4, Cdk6 and p16(INK4a), p27(KIP1) were detected with immunohistochemistry (IHC) SABC method and RT-PCR. RESULTS: After WKN intervention, the cancer cells were constrained in stage G0/G1, unable or retardatory to enter stage G (namely, the DNA synthesis stage). IHC examination showed the grey scale values of CyclinD2, CyclinE, Cdk2, Cdk4 and Cdk6 were higher, and that of p27(KIP1) and p16(INK4a) were lower in cells after moderate/high dosage WKN intervention than those in the blank control (P < 0.01); RT-PCR showed the OPTD values of CyclinD2, CyclinE, Cdk2, Cdk4 and Cdk6 were lower, and that of p27(KIP1) and p16(INK4a) were higher in cells after moderate/high dosage WKN intervention than those in the blank control (P < 0.01). CONCLUSION: WKN can inhibit the expressions of cell cycle promoting related factors of gastric cancer cell, including CyclinD2, CyclinE, Cdk2, Cdk4 and Cdk6, also enhance the expressions of cell cycle inhibiting factors of gastric cancer cell, as p27(KIP1) and p16(INK4a), which may be the mechanism of action of the remedy in preventing the growth of gastric cancer cells.
OBJECTIVE: To study the effect of Weikangning (WKN) containing serum on growth and cell cycle regulators of gastric cancer cell. METHODS: WKN containing drug serum was prepared by gastric perfusion of WKN in different dosages to SD rats. Gastric cancer cells were cultured in medium contained the drug serum with different concentrations of WKN. The change of cell cycle was detected by flow cytometry, and the mRNA and protein expressions of CyclinD2, CyclinE, Cyclin-dependant kinase2 (CdK2), Cdk4, Cdk6 and p16(INK4a), p27(KIP1) were detected with immunohistochemistry (IHC) SABC method and RT-PCR. RESULTS: After WKN intervention, the cancer cells were constrained in stage G0/G1, unable or retardatory to enter stage G (namely, the DNA synthesis stage). IHC examination showed the grey scale values of CyclinD2, CyclinE, Cdk2, Cdk4 and Cdk6 were higher, and that of p27(KIP1) and p16(INK4a) were lower in cells after moderate/high dosage WKN intervention than those in the blank control (P < 0.01); RT-PCR showed the OPTD values of CyclinD2, CyclinE, Cdk2, Cdk4 and Cdk6 were lower, and that of p27(KIP1) and p16(INK4a) were higher in cells after moderate/high dosage WKN intervention than those in the blank control (P < 0.01). CONCLUSION: WKN can inhibit the expressions of cell cycle promoting related factors of gastric cancer cell, including CyclinD2, CyclinE, Cdk2, Cdk4 and Cdk6, also enhance the expressions of cell cycle inhibiting factors of gastric cancer cell, as p27(KIP1) and p16(INK4a), which may be the mechanism of action of the remedy in preventing the growth of gastric cancer cells.