Expression of miR-19a in gastric cancer tissues and serum and its effect on proliferation and invasion ability of human gastric cancer AGS cells
10.13200/j.cnki.cjb.004406
- VernacularTitle:miR-19a在胃癌组织和血清中的表达及其对人胃癌AGS细胞增殖和侵袭能力的影响
- Author:
SHAN Biao
- Publication Type:Journal Article
- From:
Chinese Journal of Biologicals
2025;38(1):34-41+47
- CountryChina
- Language:Chinese
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Abstract:
Objective To investigate the expression of micro RNA-19a(miR-19a) in gastric cancer tissues and serum and its effect on the proliferation, invasion and regulation of thrombospondin 1(THBS1) expression in human gastric cancer AGS cells.Methods Real-time fluorescence quantitative PCR(RT-qPCR) was used to measure the expression of miR-19a in 22 gastric cancer tissues, corresponding adjacent noncancerous tissues, serum, and the serum of 22 healthy individuals. Then,the human gastric cancer AGS cells were cultured in vitro and divided into control, inhibitor NC, mimics NC, miR-19a inhibitor, miR-19a mimics, miR-19a inhibitor + si-NC and miR-19a inhibitor + si-THBS1 groups, three repeated wells for each group. The control group was without intervention, and the other groups were transfected with inhibitor NC, mimics NC, miR-19a inhibitor, miR-19a mimics, miR-19a inhibitor + si-NC and miR-19a inhibitor + si-THBS1 by using Lipofectamine~(TM)2000 liposomes respectively. After 24 h of transfection, each group was detected for the miR-19a expression, cell viability, proliferation rate, invasion number and the related proteins expression by RT-qPCR, cell counting kit-8(CCK-8), 5-ethynyl-2'-deoxyuridine(EdU), Transwell chamber and Western blot, separately.Results The expression level of miR-19a in gastric cancer tissues was significantly higher than that in corresponding adjacent tissues(t = 5. 061, P < 0. 001), and the expression level of miR-19a in serum of patients with gastric cancer was significantly higher than that of healthy people(t = 6. 299, P < 0.001). In vitro, the expression level of miR-19a in AGS cells in the miR-19a inhibitor group was significantly lower than that in the inhibitor NC group(t = 11. 120, P < 0. 001), the expression level of miR-19a in the miR-19a mimics group was significantly higher than that in the mimics NC group(t = 11. 637, P < 0. 001), and the mRNA and protein expression levels of THBS1 in the miR-19a inhibitor + si-THBS1 group were significantly lower than those in the miR-19a inhibitor + si-NC group(t = 10. 070 and 13. 164, P = 0. 001 and < 0. 001, respectively). Compared with the inhibitor NC group, the AGS cell viability, proliferation rate, invasion number, and the protein expression levels of Cyclin D1 and proliferating cell nuclear antigen(PCNA) in the miR-19a inhibitor group significantly decreased(t = 16. 679, 10. 072, 17. 737, 11. 173, and 11. 549,respectively, each P < 0. 001), and the THBS1 protein expression level increased significantly(t = 16. 774, P < 0. 001).Compared with the mimics NC group, the AGS cells in the miR-19a mimics group had significantly higher viability, proliferation rate, invasion number, and the expression levels of Cyclin D1 and PCNA proteins(t = 15. 004, 22. 746, 13. 349, 11. 484,and 19. 022, respectively, each P < 0. 001), while the expression level of THBS1 protein significantly decreased(t = 16. 384,P < 0. 001). Compared with miR-19a inhibitor + si-NC group, the viability, proliferation rate, invasion number, and the protein expression levels of Cyclin D1 and PCNA of AGS cells in miR-19a inhibitor + si-THBS1 group significantly increased(t = 13. 180, 8. 669, 14. 493, 6. 828, and 13. 587, respectively, each P < 0. 001), while THBS1 protein expression level significantly decreased(t = 14. 824, P < 0. 001).Conclusion The miR-19a is highly expressed in gastric cancer tissues and serum of patients with gastric cancer, and knockdown of miR-19a may inhibit the proliferation and invasion of human gastric cancer AGS cells by activating the expression of THBS1.
