Zhang L X, Jin W, Zheng J, Dai Y X, Song Y, Ni H B, Jiang J, Liang W B
Bratisl Lek Listy. 2018;119(1):17-21. doi: 10.4149/BLL_2018_004.
To evaluate the correlation between miRNA-375 and cell proliferation and apoptosis in glioma cancer cell.
Collecting 30 cases of glioma cancer patients and 30 cases of cerebral infarction patients. The miRNA-375 and CTGF protein expressions were evaluated by ISH and IHC methods. In the cell experiment, the U87 cells were divided into 3 groups: NC group (the cells were treated with normal method); BL group (the cells were transfected with empty vector) and miRNA group (the cells were transfected with miRNA-375). The U87 cell proliferation and apoptosis rates and cell cycle of the different groups were measured by MTT and flow cytometry. The relative proteins (CTGF, EGFR, AKT, Erk and P21) expressions were measured by WB assay.
The miRNA-375 and CTGF expressions of glioma cancer tissues were significantly different compared with those of no-cancer tissues (p < 0.05, respectively). In the cell experiments, the cell proliferation of miRNA group was significantly decreased compared with that of NC group (p < 0.05); the cell apoptosis and G1 phase rate of miRNA group was significantly decreased compared with NC group (p < 0.05, respectively). Depending on the WB assay, the CTGF, EGFR, AKT, Erk and P21 proteins expressions of miRNA group were significantly different compared with proteins expressions of NC group (p < 0.05, respectively).
miRNA-375 over-expression suppresses glioma cancer cells development via CTGF-EGFR pathway (Fig. 3, Ref. 30).
评估miRNA - 375与胶质瘤癌细胞增殖和凋亡之间的相关性。
收集30例胶质瘤患者和30例脑梗死患者。采用原位杂交(ISH)和免疫组化(IHC)方法评估miRNA - 375和结缔组织生长因子(CTGF)蛋白表达。在细胞实验中,将U87细胞分为3组:NC组(细胞采用常规方法处理);BL组(细胞转染空载体)和miRNA组(细胞转染miRNA - 375)。采用MTT法和流式细胞术检测不同组U87细胞的增殖率、凋亡率及细胞周期。采用蛋白质免疫印迹法(WB)检测相关蛋白(CTGF、表皮生长因子受体(EGFR)、蛋白激酶B(AKT)、细胞外信号调节激酶(Erk)和P21)的表达。
与非癌组织相比,胶质瘤癌组织中miRNA - 375和CTGF的表达有显著差异(分别为p < 0.05)。在细胞实验中,miRNA组的细胞增殖与NC组相比显著降低(p < 0.05);miRNA组的细胞凋亡率和G1期比例与NC组相比显著降低(分别为p < 0.05)。根据WB检测结果,miRNA组的CTGF、EGFR、AKT、Erk和P21蛋白表达与NC组蛋白表达相比有显著差异(分别为p < 0.05)。
miRNA - 375过表达通过CTGF - EGFR途径抑制胶质瘤癌细胞的发展(图3,参考文献30)。