[1]桂馥,吴宏禧,游志鹏,等.miR-21对视网膜母细胞瘤细胞增殖的影响及作用机制[J].眼科新进展,2020,40(7):630-633.[doi:10.13389/j.cnki.rao.2020.0144]
　GUI Fu,WU Hongxi,YOU Zhipeng,et al.Effects of MiR-21 on the proliferation of retinoblastoma and its mechanism[J].Recent Advances in Ophthalmology,2020,40(7):630-633.[doi:10.13389/j.cnki.rao.2020.0144]
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miR-21对视网膜母细胞瘤细胞增殖的影响及作用机制/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
40卷
期数:
2020年7期
页码:
630-633
栏目:
实验研究
出版日期:
2020-07-05

文章信息/Info

Title:
Effects of MiR-21 on the proliferation of retinoblastoma and its mechanism
作者:
桂馥; 吴宏禧; 游志鹏; 章余兰
330006　江西省南昌市，南昌大学第二附属医院眼科
Author(s):
GUI Fu;  WU Hongxi;  YOU Zhipeng;  ZHANG Yulan
Department of Ophthalmology, the Second Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province,
关键词:
miR-21; 视网膜母细胞瘤; 细胞增殖; 凋亡
Keywords:
miR-21; retinoblastoma; cell proliferation; apoptosis
分类号:
R774
DOI:
10.13389/j.cnki.rao.2020.0144
文献标志码:
A
摘要:
目的　探讨MicroRNA-21（miR-21）对视网膜母细胞瘤(RB)细胞增殖的影响及作用机制。方法　采用Real-time PCR技术检测miR-21在正常视网膜组织和确诊RB组织中的表达情况；然后在转染的基础上运用MTT检查RB细胞存活率、流式细胞仪检测细胞凋亡率。Western blot法检测与细胞凋亡相关蛋白PDCD4、Bax、Bcl-2的表达。结果　与正常视网膜组织miR-21表达 （0.703±0.071）相比，RB组织miR-21为高表达（2.214±0.162），差异有统计学意义（P<0.01）。在Weri-Rb-1细胞中，与NC组(2.245±0.213)相比，miR-21抑制剂转染后明显降低了miR-21的表达水平，miR-21 inhibitor组为0.683±0.075,差异有统计学意义 （P<0.01）。两组细胞转染后24 h、48 h、72 h、96 h，MTT测定法检测细胞活力结果显示：两组24 h的A值比较，差异无统计学意义 （P>0.05），miR-21 inhibitor 组在 48 h、72 h、 96 h的A值均低于 NC 组，差异均有统计学意义 （均为P<0.01）。流式细胞术检测结果显示：NC组凋亡细胞在总细胞中百分比为（3.045±0.301）%和（4.832±0.493）%，miR-21 inhibitor组凋亡细胞在总细胞中百分比为（2.593±0.257）%和（40.167±4.014）%，miR-21 inhibitor组Weri-Rb-1细胞的凋亡率明显高于miR-21 NC组(P<0.01)。Western blot检测结果显示：NC组PDCD4表达（0.192±0.045）相比miR-21 inhibitor组（0.683±0.091）表达明显减少，NC组Bax的蛋白表达水平（0.143±0.036）相比miR-21 inhibitor组（1.192±0.054）也明显减少，差异均有统计学意义（P<0.01)，NC组Bcl-2蛋白表达（0.864±0.038）相比miR-21 inhibitor组（0.257±0.026）明显增多，差异有统计学意义（P<0.05)。结论　miR-21是RB的促癌基因，miR-21抑制剂可以通过降低miR-21表达抑制肿瘤细胞的增殖、促进细胞凋亡，这一过程与PDCD4、Bax、Bcl-2等凋亡相关蛋白有关。
Abstract:
Objective　To investigate the effect of MicroRNA-21 (miRNA-21) on retinoblastoma (RB) cell proliferation and its possible mechanisms. Methods　Expression of miR-21 in RB tissue and human normal tissue was measured Real-time PCR. RB cells were transfected with miR-21 control or miR-21 inhibitor via LipofectaminTM 2000. Cell viability was analyzed by MTT assay. Flow cytometry with Annexin V-FITC/PI reagent was used to detect cell apoptosis. The protein levels of Bax, Bcl-2and PDCD4 were examined by Western blot. Results　miR-21 was overexpressed in RB tissues (2.214±0.162) when compared with normal retinal tissues (0.703±0.071) with significant difference (P<0.01). When compared with normal control (NC) group (2.245±0.213), in Weri-Rb-1 cells, miR-21 inhibitor transfection suppressed the expression of miR-21 in miR-21 inhibitor group (0.683±0.073) (P<0.01). In addition, 24 hours, 48 hours, 72 hours, 96 hours after transfection of the two groups of cells, the results of the viability of the cells via MTT assay showed that there was no statistically significant difference in the A value between the two groups at 24 hours (P>0.05), A values of miR-21 inhibitor group at 48 h, 72 h, 96 h were lower than those of the NC group, and the differences are statistically significant (all P<0. 01). The results of flow cytometry showed that the percentage of apoptotic cells in the NC group was (3.045 ± 0.301)% and (4.832 ± 0.493)%, and (2.593 ± 0.257)% and (40.167 ± 4.014)% in the miR-21 inhibitor group. The apoptosis rate of the Weir-Rb-1 cells in the miR-21 inhibitor group was significantly higher than that in the miR-21 NC group (P<0.01). The Western blot results showed that the expression of PDCD4 in the NC group (0.192 ± 0.045) was significantly reduced compared to the miR-21 inhibitor group (0.683 ± 0.091), and the expression level of the Bax protein in the NC group was 0.143 ± 0.036, which was significantly reduced compared to the miR-21 inhibitor group (1.192±0.054), and the difference was also statistically significant (P<0.01). The expression of Bcl-2 protein in the NC group (0.864±0.038) was significantly enhanced when compared with the miR-21 inhibitor group (0.257 ± 0.026), and the difference was statistically significant (P<0.05). Conclusion　miR-21 promotes carcinogenesis in the development and progression of RB. miR-21 inhibitor can inhibit the proliferation of Weri-RB-1 cells and increase the apoptosis of Weri-RB-1 cells by decreasing miR-21 expression. This process is related to apoptosis-related proteins such as PDCD4, Bax, and Bcl-2.

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更新日期/Last Update: 2020-07-05