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

[目的]探讨环状RNA(circRNA)circXPO1通过mi R-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(q RT-PCR)和Western blot测定circXPO1、mi R-5195-3p、RTKN mRNA和RTKN蛋白的表达。SW620细胞分为si-NC组、si-circXPO1组、mi R-NC组、mi R-5195-3p组、si-circXPO1+anti-mi R-5195-3p组、si-circXPO1+pcDNA-RTKN组,依次转染si RNA-NC(si-NC)、si RNA-circXPO1(si-circXPO1)、mimic NC(mi R-NC)、mi R-5195-3p mimic、si-circXPO1、mi R-5195-3p inhibitor(anti-mi R-5195-3p)、si-circXPO1和pc DNA-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mi R-5195-3p、mi R-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 m RNA(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,mi R-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±0.03vs 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mi R-NC组增加,差异有统计学意义(P<0.05);si-circXPO1+anti-mi R-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mi R-5195-3p/RTKN轴促进结直肠癌SW620细胞增殖,和抑制其凋亡。

[Objective]To explore the mechanism of circular RNA(circ RNA)circ XPO1 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-circ XPO1 group,mi R-NC group,mi R-5195-3 p group,si-circ XPO1+anti-mi R-5195-3 p and si-circ XPO1+pc DNA-RTKN group,which were transfected with si RNA-NC(si-NC),si RNA-circ XPO1(si-circ XPO1),mimic NC(mi R-NC),mi R-5195-3 p mimic,si-circ XPO1 and mi R-5195-3 p inhibitor(anti-mi R-5195-3 p),si-circ XPO1 and pc DNA-RTKN,respectively.Real-time quantitative polymerase chain reaction(q RTPCR)and Western blot were performed to determine the expression of circ XPO1,mi R-5195-3 p,RTKN m RNA 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 circ XPO1 with mi R-5195-3 p,mi R-5195-3 p and RTKN.[Results]Compared with adjacent tissues,the expression levels of circ XPO1(4.55±0.30 vs 4.55±0.30),RTKN m RNA(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,mi R-5195-3 p(0.34±0.03 vs 1.00±0.06)expression level decreased(all P<0.05).The expression level of circ XPO1 and cell viability(0.37±0.03 vs 0.72±0.05)in colorectal cancer SW620 cells of the si-circ XPO1 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).Circ XPO1 targets the expression of mi R-5195-3 p,and mi R-5195-3 p 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 mi R-5195-3 p group were lower than those in the mi R-NC group,and the apoptosis rate(21.01%±2.11%vs 7.17%±0.54%)was higher than that in the mi R-NC group(all P<0.05).The expression level of RTKN protein and cell activity(0.34±0.03 and 0.38±0.04 vs0.17±0.02)of the si-circ XPO1+anti-mi R-5195-3 p group and si-circ XPO1+pc DNA-RTKN group were higher than those of the sicirc XPO1 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 sicirc XPO1 group(all P<0.05).[Conclusion]Circ XPO1 promotes the proliferation of colorectal cancer SW620 cells and inhibits cell apoptosis through the mi R-5195-3 p/RTKN axis.