鲁 斌,黄晓宇,夏秀梅.MiR-184在人胃癌组织中的表达及对胃癌细胞凋亡的影响[J].肿瘤学杂志,2021,27(2):130-135. |
MiR-184在人胃癌组织中的表达及对胃癌细胞凋亡的影响 |
Expression of MiR-184 in Gastric Cancer and Its Effect on Apoptosis of Gastric Cancer Cells |
投稿时间:2019-10-16 |
DOI:10.11735/j.issn.1671-170X.2021.02.B008 |
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中文关键词: 胃肿瘤 miRNA 转染 凋亡 侵袭 |
英文关键词:gastric cancer miRNA transfection apoptosis invasion |
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中文摘要: |
摘 要:[目的] 探讨miR-184在人胃癌组织中的表达及对胃癌细胞凋亡的影响。[方法] 使用Real-time PCR检测miR-184表达,SGC-7901细胞转染,MTT法检测细胞增殖情况,Hoechst33342染色观察细胞自噬、凋亡情况,Transwell法检测细胞侵袭能力应用,流式细胞术检测细胞凋亡情况,Western blot检测各组细胞NF-κB、LC3Ⅱ、Beclin1蛋白表达。[结果](1)胃癌组织的miR-184表达水平显著低于正常胃组织(1.74±0.12 vs 2.51±0.41,t=12.745,P<0.001)。(2)转染1d后,miR-184 mimics组的miR-184表达水平(6.02±1.15)显著高于NC组(2.15±0.37)(t=22.652,P<0.001),miR-184 inhibitors组表达水平(0.52±0.09)显著低于NC组(t=30.268,P<0.001)。(3)24~72h内,miR-184 inhibitors组和NC组的细胞增殖能力显著性高于miR-184 mimics组(P<0.05)。(4)MiR-184 mimics组(45.39%±1.74%)侵袭率显著低于NC组(52.05%±2.03%)(t=17.614,P<0.001),而miR-184 inhibitors组(94.51%±3.08%)侵袭率显著高于NC组(t=22.652,P<0.001)。(5)MiR-184 mimics组(72.32%±22.82%)细胞凋亡率显著高于NC组(44.07%±16.23%)(t=7.133,P<0.001),而miR-184 inhibitors组(35.31%±11.29%)凋亡率显著低于NC组细胞(t=3.133,P=0.001)。(6)MiR-184 mimics组可见大量自噬泡,NC组自噬泡可见部分,miR-184 inhibitors组仅可见极少量自噬泡。(7)MiR-184 mimics组NF-κB、LC3Ⅱ、Beclin1蛋白表达水平显著高于NC组(t=14.055、14.486、28.450,P均<0.001),miR-184 inhibitors组NF-κB、LC3Ⅱ、Beclin1蛋白表达水平显著低于NC组(t=15.423、10.354、39.836,P均<0.001)。[结论] MiR-184可在一定程度上抑制胃癌细胞的侵袭和增殖能力,其主要作用机制是诱导胃癌细胞的自噬与凋亡。 |
英文摘要: |
Abstract:[Objective] To investigate the expression of miR-184 in gastric cancer and its effect on apoptosis of gastric cancer cells. [Methods] MiR-184 expression was detected by real-time RT-PCR. Human gastric cancer SGC-7901 cells were transfected with miR-184. The cell proliferation was detected by MTT method,autophagy and apoptosis were observed by Hoechst 33342 staining,cell invasion was detected by Transwell method,cell apoptosis was detected by flow cytometry,the expression of NF-κB,LC3 Ⅱ and Beclin1 protein were detected by Western blot. [Results] The expression level of miR-184 in gastric cancer was significantly lower than that in normal gastric tissue(1.74±0.12 vs 2.51±0.41,t=12.745,P<0.001). The expression level of miR-184 in miR-184 mimics group was significantly higher than that in NC group(6.02±1.15 vs 2.15±0.37,t=22.652,P<0.001),and the expression level of miR-184 inhibitor group(0.52±0.09) was significantly lower than that in NC group(t=30.268,P<0.001). The proliferation ability of miR-184 inhibitor group and NC group was significantly higher than that of miR-184 mimics group(P<0.05). The invasion rate of miR-184 mimics group(45.39%±1.74%) was significantly lower than that of NC group(52.05%±2.03%)(t=17.614,P<0.001),while that of miR-184 inhibitor group(94.51%±3.08%) was significantly higher than that of NC group(t=22.652,P<0.001). The apoptosis rate of miR-184 mimics group(72.32%±22.82%) was significantly higher than that of NC group(44.07%±16.23%)(t=7.133,P<0.001),while that of miR-184 inhibitor group(35.31%±11.29%) was significantly lower than that of NC group(t=3.133,P=0.001). A large number of autophagic vesicles were observed in the miR-184 mimics group,but not seen in the NC group,only a very small number were seen in the miR-184 inhibitor group. The expression levels of NF-κB,LC3Ⅱ,Beclin1 protein in miR-184 mimics group were significantly higher than those in NC group(t=14.055,14.486,28.450,all P<0.001),and the expression of those proteins in miR-184 inhibitor group were significantly lower than those in the NC group(t=15.423,10.354,39.836,all P<0.001). [Conclusion] miR-184 can inhibit the proliferation and invasion of gastric cancer cells to a certain extent,and inducing autophagy and apoptosis of gastric cancer cells may be the main mechanism. |
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