1.Department of GastrointestinalSurgery, the Second Affiliated Hospital & Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou325027, China; 2.Department of Operating Room, the Second Affiliated Hospital & Yuying Children’s Hospital ofWenzhou Medical University, Wenzhou 325027, China; 3.Department of Gastroenterology, the Second AffiliatedHospital & Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou 325027, China
CHEN Jian,LI Liyi,ZHUGE Linmin, et al. Effect of Hsa-miR-145 targeting replication protein A1 on proliferation of gastric cancer[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2023, 53(11): 917-922.
Abstract：Objective: To investigate the effect of miRNA targeting replication protein A1(RPA1) gene in gastric cancer cells on tumor malignant biological behavior and its mechanism. Methods: Bioinformatics analysis was used to compare the expression of RPA1 gene in cancer and adjacent tissues and its correlation with the prognosis of gastric cancer patients. RNA pulldown method was used to detect the miRNA targeting the 3’UTR region of RPA1 gene in gastric cancer cells (SGC-7901); SGC-7901 cells with high expression of miR-145 were constructed. Cell proliferation was detected by MTT staining, and cell cycle phase was analyzed by flow cytometry; The targeting relationship between miR-145 and RPA1 was verified by luciferase experiment, which was further confirmed by Western blot. Results: Compared with the adjacent tissues, the expression of RPA1 gene was significantly up-regulated in gastric cancer tissues. The high expression of RPA1 gene in gastric cancer tissues was associated with worse prognosis of patients. RNA pulldown was used to detect miRNA targeting the 3’UTR region of RPA1 gene in gastric cancer cell SGC-7901. The results showed that miR-145 had the highest binding with the 3’UTR region of RPA1 gene. Then, miR-145 was selected for further research. The results of luciferase reporter gene experiment showed that the luciferase activity was lower than that in the control miRNA group (P<0.05); the Western blot results showed that the expression of RPA1 in gastric cancer cells in miR-145group was lower than that in the control miRNA group (P<0.05). MTT results showed that the proliferation ability of gastric cancer cells in miR-145 group was lower than that in control miRNA group (P<0.05); Flow cytometry showed that the G2/M phase arrest of gastric cancer cells in miR-45 group increased (P<0.05). Conclusion: The overexpression of miR-145 can significantly inhibit the proliferation of gastric cancer cell SGC-7901, and its mechanism may be related to down-regulation of RPA1 expression which blocks the cell cycle.
