Expression of miRNA-372-3p in gastric cancer tissues and its effect on the biological function of gastric cancer cells
10.3760/cma.j.cn115355-20200611-00315
- VernacularTitle:miRNA-372-3p在胃癌组织中的表达及其对胃癌细胞生物学功能的影响
- Author:
Zongliang GUO
1
;
Feng LI
Author Information
1. 山西省肿瘤医院普外科，太原 030013
- From:
Cancer Research and Clinic
2020;32(8):529-534
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the expression of miRNA-372-3p (miR-372-3p) in gastric cancer tissues and cells and the regulation of RAB22A protein as well as its effect of miR-372-3p on the biological function of gastric cancer cells.Methods:The expression levels of miR-372-3p in cancer tissues of 70 patients clinically diagnosed with gastric cancer and the pericarcinomatous tissues were detected by using real-time quantitative polymerase chain reaction (RT-PCR) from Shanxi Provincial Cancer Hospital between December 2018 and December 2019. miR-372-3p inhibitor was transfected in gastric cancer MGC-803 and SGC-7901 cells, and miR-372-3p NC (empty vector) was used as the control. The colony formation, proliferation and apoptosis of gastric cancer cells were detected by using the clone formation assay, CCK-8 method and flow cytometry, respectively. Dual-luciferase reporter gene assay was used to detect the expression correlation between miR-372-3p and RAB22A of MGC-803 cells. miRNA-21 (miR-21) was treated as the negative control, and Western blot was used to detect the expression of protein RAB22A in MGC-803 and SGC-7901 cells when miRNA-21 (miR-21) was treated as the negative control.Results:RT-PCR results showed that the mRNA relative expression level of miR-372-3p was up-regulated in gastric cancer tissues compared to their adjacent normal cancer tissues, and the difference was statistically significant (0.51±0.37 vs. 0.77±0.48, t = 1.98, P = 0.005). There were statistically significant differences in mRNA expression level of miR-372-3p in gastric cancer tissues with different tumor diameter and pathological grade (all P < 0.05). The assay in vitro showed that the low expression of miR-372-3p could inhibit the clone formation of MGC-803 and SGC-7901 cells [(211±4) vs. (410±5), t = 2.78, P = 0.001; (244±8) vs. (423±7), t = 2.76, P = 0.001], cell proliferation activity (absorbance value of MGC-803 cells for 48 h: 0.39±0.06 vs. 0.57±0.03, t = 3.18, P = 0.01; absorbance value of MGC-803 cells for 72 h: 0.50±0.05 vs. 0.81±0.06, t = 2.78, P < 0.01; absorbance value of SGC-7901 cells for 72 h: 0.50±0.09 vs. 0.79±0.09, t = 2.77, P = 0.01) and increase the early apoptosis rate of MGC-803 cells [(25.19±0.26) vs. (20.02±0.04), t = 4.30, P < 0.05]. Dual-luciferase reporter gene assay found that compared with the cotransfection of miR-372-3p NC and RAB22A wild type gene, the luciferase activity of MGC-803 cells was decreased after the cotransfection of miR-372-3p inhibitor and RAB22A wild type gene, and the difference was statistically significant [(1.00±0.04) vs. (0.53±0.06), t = 3.18, P = 0.01]. Western blot results showed that knockdown of miR-372-3p could inhibit the expressions of MGC-803, SGC-7901 cells and RAB22A protein. Conclusion:The expression of miR-372-3p in gastric cancer tissues is up-regulated, and miR-372-3p can promote the expression of RAB22A and regulate the occurrence and development of gastric cancer. It may be a potential therapeutic target.
