RAB10对胰腺癌细胞生物学功能的影响及临床意义

Effect of RAB10 on biological function of pancreatic cancer cells and its clinical significance

目的研究RAS癌基因家族成员RAB10在胰腺癌中的表达情况及其对人胰腺癌细胞系SW1990细胞的增殖、迁移、侵袭和凋亡的影响。方法利用癌症基因数据库GEPIA和TCGA中胰腺癌的数据分析RAB10 mRNA在胰腺癌组织中的表达;Cox回归分析RAB10 mRNA表达水平与胰腺癌患者预后关系;靶向RAB10的小干扰RNA(RAB10-siRNA)作为沉默组、构建过表达RAB10质粒(RAB10-OE)作为过表达组;使用Q-PCR检测沉默及过表达效果;Western blot实验检测蛋白表达水平;EdU细胞增殖实验、划痕实验、Transwell实验、流式细胞实验分别分析RAB10对SW1990增殖、迁移、侵袭和凋亡的影响。结果胰腺癌组织中RAB10 mRNA的表达量明显高于正常胰腺组织(P<0.05);EdU细胞增殖实验结果显示,RAB10-OE组的SW1990细胞的增殖率明显高于对照组,RAB10-siRNA组SW1990细胞的增殖率低于对照组(P<0.05);Transwell实验和划痕实验结果显示,RAB10-OE组细胞侵袭率及迁移率均高于对照组,RAB10-siRNA组细胞迁移率及侵袭率均低于对照组(P<0.05);流式细胞实验结果显示,RAB10-OE组细胞凋亡率低于对照组,RAB10-siRNA组细胞凋亡率高于对照组(P<0.05);RAB10高表达组生存时间低于RAB10低表达组(P<0.05);单因素Cox回归分析结果显示,临床分级、T分期、M分期及RAB10 mRNA表达水平与胰腺癌患者预后相关(P<0.05)。多因素Cox回归分析结果显示,RAB10 mRNA的表达水平为影响胰腺癌患者预后的独立危险因素(P<0.05)。结论RAB10在胰腺癌组织中高表达,是胰腺癌患者预后的独立危险因素且能够促进胰腺癌细胞的增殖、侵袭和迁移,并抑制胰腺癌细胞的凋亡。

Objective To investigate the expression of Member RAS Oncogene Family(RAB10)in pancreatic cancer(PAAD)and its effects on the proliferation,migration,invasion and apoptosis of SW1990 cells(human pancreatic cancer cells).Methods The expression of RAB10 mRNA in PAAD tissues wasanalyzed by the cancer gene database GEPIA(Gene Expression Profiling Interactive Analysis)and TCGA(The Cancer Genome Atlas).Cox regression analysis was used to detect relationship between RAB10 mRNA expression and the prognosis of pancreatic cancer patients.We targeted small interfering RNA(RAB10-siRNA)targeting RAB10 as the silence group,and constructed an overexpression plasmid(RAB10-OE)for overexpression of RAB10 as the overexpression group.The effects of silencing and overexpressionweredetected by Q-PCR;protein expression levelsweredetected by Western blot.EdUcellproliferation test,wound healing test,Transwelltestand flow cytometry test were used to determine the effects of RAB10 on the proliferation,migration,invasion and apoptosis of SW1990 pancreatic cancer cells.Results RAB10 mRNA expression in PAAD tissues was higher than that innormal pancreatic tissues(P<0.05).The results of EdUcellproliferation testshowed that the proliferation rate of SW1990 cells in the RAB10-OE group was higher thanthat in the control group,and the proliferation rate of SW1990 cells in the RAB10-siRNA group was lower than that inthe control group(P<0.05).The results of the Transwell test and wound healing test showed that the invasion rate and mobility rate of RAB10-OE group were higher thanthose of the control group,and the mobility and invasion rate of RAB10-siRNA group were lower than those of the control group(P<0.05).The results of flow cytometry test showed that the apoptosis rate was lower in the RAB10-OE group than the control group,and the apoptosis rate in the RAB10-siRNA group was higher than the control group(P<0.05).The median survival time of RAB10 high expression group was significantly lower than that of RAB10 low expression group(P<0.05).Cox regression analysis showed that clinical grade,T stage,M stage and RAB10 mRNA expression were related to survival and prognosis of pancreatic cancerpatients(P<0.05).Multivariate Cox regression analysis showed that the expression level of RAB10 mRNA was the independent risk factor affecting the prognosis of pancreatic cancer patients(P<0.05).Conclusion RAB10 is highly expressed in PAAD tissues and RAB10 can promote the proliferation of pancreatic cancer cells,accelerate the ability to invade and migrate,and inhibit the apoptosis of pancreatic cancer cells.RAB10 is an independent risk factor for survival prognosis in patients with pancreatic cancer.