Na Deng1, Di Lei1, Jinfa Huang1, Zhuanhong Yang2, Cuifang Fan3, Suqing Wang4. 1. Department of Obstetrics and Gynecology, Renmin Hospital of Wuhan University, Wuhan 430060, China. 2. Department of Nutrition and Food Hygiene, School of Health Science, Wuhan University, 185, Donghu Rd, Wuhan, Hubei 430071, China. 3. Department of Obstetrics and Gynecology, Renmin Hospital of Wuhan University, Wuhan 430060, China. Electronic address: fcf728@163.com. 4. Department of Nutrition and Food Hygiene, School of Health Science, Wuhan University, 185, Donghu Rd, Wuhan, Hubei 430071, China. Electronic address: swang2099@whu.edu.cn.
Abstract
OBJECTIVES: To identify circRNA expression profiles in the placentae of severe preeclampsia (SPE) and normal pregnant (NP) women. METHODS: Placental samples were collected from six paired SPE and NP women. CircRNA expression profiles were identified by RNA-Seq and validated in another 30 SPE and NP samples by qRT-PCR. Several bioinformatic tools were utilised to analyse the potential function of differentially expressed circRNAs (DE-circRNAs) and to predict target microRNAs (miRNAs) and proteins. Furthermore, immunohistochemistry, Western blotting and RNA binding protein immunoprecipitation (RIP) were conducted to confirm the interaction between the circRNA and protein. RESULTS: In total, 18,631 circRNAs were detected. Among them, 180 circRNAs were differentially expressed, including 94 upregulated and 86 downregulated circRNAs. Seven DE-circRNAs were selected for validation, and the results of six circRNAs (hsa_circ_0007611, hsa_circ_0011460, hsa_circ_0002888, and hsa_circ_0007445, hsa_circ_0017068, hsa_circ_0012737) were consistent with the sequencing results. Bioinformatics analyses of DE-circRNAs revealed that most of them are involved in vasodilation, regulation of blood vessel size, protein transport and localization, and pathways in cancer. In addition to the mRNA expression profile, it was interesting to find that the hsa_circ_0011460 target gene solute carrier organic anion transporter family member 2A1 (SLCO2A1, PGT) was also significantly increased in SPE placentae. Immunohistochemistry, Western blotting and qRT-PCR validated the expression and distribution of PGT. Finally, RIP of HTR-8/SVneo cells confirmed that hsa_circ_0011460 targets PGT directly. CONCLUSIONS: A total of 180 DE-circRNAs were detected. One crucial circRNA, hsa_circ_0011460, was shown to interact directly with its host gene PGT. These findings indicated that hsa_circ_0011460 may serve as a potential therapeutic target for patients with SPE.
OBJECTIVES: To identify circRNA expression profiles in the placentae of severe preeclampsia (SPE) and normal pregnant (NP) women. METHODS: Placental samples were collected from six paired SPE and NP women. CircRNA expression profiles were identified by RNA-Seq and validated in another 30 SPE and NP samples by qRT-PCR. Several bioinformatic tools were utilised to analyse the potential function of differentially expressed circRNAs (DE-circRNAs) and to predict target microRNAs (miRNAs) and proteins. Furthermore, immunohistochemistry, Western blotting and RNA binding protein immunoprecipitation (RIP) were conducted to confirm the interaction between the circRNA and protein. RESULTS: In total, 18,631 circRNAs were detected. Among them, 180 circRNAs were differentially expressed, including 94 upregulated and 86 downregulated circRNAs. Seven DE-circRNAs were selected for validation, and the results of six circRNAs (hsa_circ_0007611, hsa_circ_0011460, hsa_circ_0002888, and hsa_circ_0007445, hsa_circ_0017068, hsa_circ_0012737) were consistent with the sequencing results. Bioinformatics analyses of DE-circRNAs revealed that most of them are involved in vasodilation, regulation of blood vessel size, protein transport and localization, and pathways in cancer. In addition to the mRNA expression profile, it was interesting to find that the hsa_circ_0011460 target gene solute carrier organic anion transporter family member 2A1 (SLCO2A1, PGT) was also significantly increased in SPE placentae. Immunohistochemistry, Western blotting and qRT-PCR validated the expression and distribution of PGT. Finally, RIP of HTR-8/SVneo cells confirmed that hsa_circ_0011460 targets PGT directly. CONCLUSIONS: A total of 180 DE-circRNAs were detected. One crucial circRNA, hsa_circ_0011460, was shown to interact directly with its host gene PGT. These findings indicated that hsa_circ_0011460 may serve as a potential therapeutic target for patients with SPE.
Authors: Aleksandra Lipka; Jan Pawel Jastrzebski; Lukasz Paukszto; Karol Gustaw Makowczenko; Elzbieta Lopienska-Biernat; Marek Gowkielewicz; Ewa Lepiarczyk; Marta Wiszpolska; Mariusz Krzysztof Majewski; Marta Majewska Journal: Cells Date: 2021-04-16 Impact factor: 6.600