| 李 洁,黎钟燕,辜金莲.CircXPO1/miR-5195-3p/RTKN轴调控结直肠癌SW620细胞增殖和凋亡的机制研究[J].肿瘤学杂志,2021,27(8):615-621. |
| CircXPO1/miR-5195-3p/RTKN轴调控结直肠癌SW620细胞增殖和凋亡的机制研究 |
| CircXPO1/miR-5195-3p/RTKN Axis in Regulating the Proliferation and Apoptosis of Colorectal Cancer Cells |
| 投稿时间：2021-02-25 |
| DOI：10.11735/j.issn.1671-170X.2021.08.B003 |
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| 中文关键词: 结直肠肿瘤 SW620细胞 circXPO1 miR-5195-3p RTKN 增殖 凋亡 |
| 英文关键词:colorectal cancer SW620 cells circXPO1 miR-5195-3p RTKN proliferation apoptosis |
| 基金项目:广东省科技计划项目（0445091120222076；01548280218329060） |
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| 中文摘要: |
| 摘 要：［目的］ 探讨环状RNA（circRNA）circXPO1通过miR-5195-3p/RTKN轴调控结直肠癌SW620细胞增殖和凋亡的机制。［方法］ 应用实时定量聚合酶链反应（qRT-PCR）和Western blot测定circXPO1、miR-5195-3p、RTKN mRNA和RTKN蛋白的表达。SW620细胞分为si-NC组、si-circXPO1组、miR-NC组、miR-5195-3p组、si-circXPO1+anti-miR-5195-3p组、si-circXPO1+pcDNA-RTKN组，依次转染siRNA-NC（si-NC）、siRNA-circXPO1（si-circXPO1）、mimic NC（miR-NC）、miR-5195-3p mimic、si-circXPO1、miR-5195-3p inhibitor（anti-miR-5195-3p）、si-circXPO1和pcDNA-RTKN。运用细胞计数试剂盒8（CCK-8）、流式细胞术检测SW620细胞增殖、凋亡。生物信息学分析与双荧光素酶报告实验测定circXPO1与miR-5195-3p、miR-5195-3p与RTKN的靶向关系。［结果］ 与癌旁组织比较，结直肠癌组织中circXPO1（4.55±0.30 vs 4.55±0.30）、RTKN mRNA（3.27±0.29 vs 1.00±0.04）和RTKN蛋白（0.51±0.06 vs 0.21±0.03）表达水平升高，miR-5195-3p（0.34±0.03 vs 1.00±0.06）表达水平降低，差异有统计学意义（P<0.05）。Si-circXPO1组结直肠癌SW620细胞中circXPO1表达水平、细胞活性（0.37±0.03 vs 0.72±0.05）均比si-NC组低，凋亡率（25.91%±2.51% vs 6.73%±0.65%）比si-NC组高，差异有统计学意义（P<0.05）。CircXPO1靶向调控miR-5195-3p的表达，miR-5195-3p靶向调控RTKN的表达。MiR-5195-3p组结直肠癌SW620细胞中RTKN蛋白、细胞活性（0.43±0.04 vs 0.74±0.05）比miR-NC组减少，凋亡率（21.01%±2.11% vs 7.17%±0.54%）比miR-NC组增加，差异有统计学意义（P<0.05）；si-circXPO1+anti-miR-5195-3p组和si-circXPO1+pcDNA-RTKN组RTKN蛋白表达水平、细胞活性（0.34±0.03、 0.38±0.04、0.17±0.02）均高于si-circXPO1组，凋亡率（13.83%±1.21%、10.19%±0.82%、27.53%±2.65%）均低于si-circXPO1组，差异有统计学意义（P<0.05）。 ［结论］ CircXPO1通过miR-5195-3p/RTKN轴促进结直肠癌SW620细胞增殖，和抑制其凋亡。 |
| 英文摘要: |
| Abstract： ［Objective］ To explore the mechanism of circular RNA(circRNA) circXPO1 in regulating the proliferation and apoptosis of colorectal cancer cells. ［Methods］ Surgical samples of cancer and adjacent tissue were collected from 41 patients with colorectal cancer. Human colorectal cancer SW620 cells were divided into si-NC group， si-circXPO1 group， miR-NC group， miR-5195-3p group， si-circXPO1+anti-miR-5195-3p and si-circXPO1+pcDNA-RTKN group， which were transfected with siRNA-NC(si-NC)， siRNA-circXPO1(si-circXPO1)， mimic NC(miR-NC)， miR-5195-3p mimic， si-circXPO1 and miR-5195-3p inhibitor(anti-miR-5195-3p)， si-circXPO1 and pcDNA-RTKN， respectively. Real-time quantitative polymerase chain reaction(qRT-PCR) and Western blot were performed to determine the expression of circXPO1， miR-5195-3p， RTKN mRNA and RTKN protein. Cell counting kit 8(CCK-8) and flow cytometry were used to detect the proliferation and apoptosis of SW620 cells. Bioinformatics analysis and dual luciferase report experiment were performed to determine the targeting relationship of circXPO1 with miR-5195-3p， miR-5195-3p and RTKN. ［Results］ Compared with adjacent tissues， the expression levels of circXPO1(4.55±0.30 vs 4.55±0.30)， RTKN mRNA(3.27±0.29 vs 1.00±0.04) and RTKN protein(0.51±0.06 vs 0.21±0.03) in colorectal cancer tissues increased， miR-5195-3p(0.34±0.03 vs 1.00±0.06) expression level decreased(all P<0.05). The expression level of circXPO1 and cell viability(0.37±0.03 vs 0.72±0.05) in colorectal cancer SW620 cells of the si-circXPO1 group were lower than those in the si-NC group； and the apoptosis rate(25.91%±2.51% vs 6.73%±0.65%) was higher than that in the si-NC group. The difference was statistically significant(P<0.05). CircXPO1 targets the expression of miR-5195-3p， and miR-5195-3p targets the expression of RTKN. The RTKN protein and cell activity(0.43±0.04 vs 0.74±0.05) of colorectal cancer SW620 cells in the miR-5195-3p group were lower than those in the miR-NC group， and the apoptosis rate(21.01%±2.11% vs 7.17%±0.54%) was higher than that in the miR-NC group(all P<0.05). The expression level of RTKN protein and cell activity(0.34±0.03 and 0.38±0.04 vs 0.17±0.02) of the si-circXPO1+anti-miR-5195-3p group and si-circXPO1+pcDNA-RTKN group were higher than those of the si-circXPO1 group， and the apoptosis rates(13.83%±1.21% and 10.19%±0.82% vs 27.53%±2.65%) were lower than that of the si-circXPO1 group(all P<0.05). ［Conclusion］ CircXPO1 promotes the proliferation of colorectal cancer SW620 cells and inhibits cell apoptosis through the miR-5195-3p/RTKN axis. |
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