高尔基磷蛋白3对5-氟尿嘧啶化疗耐药的人结肠癌细胞影响研究

Golgi phosphoprotein 3 gene expression promotes the resistance of colon cancer cell to 5-fluorouracil

目的探讨人结肠癌细胞中高尔基磷蛋白3(GOLPH3)基因高表达与结肠癌细胞对5-氟尿嘧啶(5-FU)的耐药的相关性。方法HT29细胞分为5组:对照组(HT29),转染组(siRNA-GOLPH3),实验组1(30μmol·L^-15-FU),实验组2(siRNA-GOLPH3+30μmol·L^-15-FU),实验组3(30μmol·L-15-FU+50μmol·L^-1PD98059)。人结肠癌细胞转染siRNA-GOLPH3,以逆转录聚合酶链反应方法检测GOLPH3基因的沉默效果,以噻唑蓝、流式细胞术检测各组细胞增殖和凋亡,以Western blot法检测GOLPH3,P糖蛋白(P-gp),ERK1/2和p ERK1/2蛋白的表达。结果对照组和转染组的GOLPH3 mRNA的蛋白相对表达量分别为1.00±0.08和0.15±0.02,差异有统计学意义(P<0.05),表明GOLPH3基因沉默效果好。实验组1、实验组2和对照组的增殖能力分别为0.80±0.10,0.24±0.09和1.05±0.14,凋亡率分别为(12.87±2.20)%,(34.23±4.76)%和(2.50±1.47)%,实验组1、实验组2的上述指标与对照组比较,差异均有统计学意义(均P<0.05)。对照组的GOLPH3、P-gp、p ERK1/2蛋白表达量分别为0.97±0.10,1.00±0.20和1.00±0.33,实验组1分别为3.63±0.57,2.42±0.26和2.66±0.44,实验组2分别为0.52±0.18,0.47±0.16和0.24±0.09,实验组1的上述指标与实验组2比较,差异均有统计学意义(均P<0.05)。实验组3的P-gp蛋白表达量(0.55±0.04)较实验组1及对照组均明显降低(P<0.05)。结论GOLPH3高表达可能参与HT29结肠癌细胞对5-FU化疗耐药,其机制可能通过激活MAPK/ERK细胞信号通路而促进HT29结肠癌细胞对5-FU耐药。沉默GOLPH3表达可一定程度上逆转其耐药。

Objective pression of Golgi phosphoprotein 3(GOLPH3)gene and resistance of HT29 colon cancer cell to 5-fluorouracil(5-FU).Methods cancer cells were divided into five groups:control group,siRNA transfection group,experimental-1 group(30μmol·L^-15-FU),experimental-2 group(siRNA-GOLPH3+30μmol·L^-15-FU)and experimental-3 group(30μmol·L^-15-FU+50μmol·L^-1 PD98059).The silencing effect of GOLPH3 was detected by reverse transcription-polymerase chain reaction,and methyl thiazolyl tetrazolium and flow cytometry were respectively used to detect the proliferation and apoptosis of HT29 cells.The expressions of GOLPH3,P-glycoprotein(P-gp),ERK1/2 and p ERK1/2 in HT29 cell were detected by Western blot.Results 1.00±0.08 and 0.15±0.02,compared with control group,the relative expression in transfection group was significantly decreased(P<0.05).It proved the silencing effect of GOLPH3 was good.The proliferation ability of experimental-1 group,experimental-2 group and control group were 0.80±0.10,0.24±0.09,1.05±0.14,the apoptosis rates were(12.87±2.20)%,(34.23±4.76)%and(2.50±1.47)%,the were significant difference in experimental-1 group and experimental-2 group compared with control group(all P<0.05).The expression of GOLPH3,P-gp and pERK1/2 of control group were 0.97±0.10,1.00±0.20 and 1.00±0.33,those in experimental-1 group were 3.63±0.57,2.42±0.26 and 2.66±0.44,and those in experimental-2 group were 0.52±0.18,0.47±0.16 and 0.24±0.09,there were significant differences between experimental-1 group and experimental-2 group(all P<0.05).The expression of P-gp in experimental-3 group was 0.55±0.04,which was significantly lower than that in experimental-1 group and control group(P<0.05).Conclusion may be involved in the resistance of HT29 colon cancer cells to 5-FU chemotherapy.The mechanism may promote the resistance of HT29 colon cancer cells to 5-FU by activating the MAPK/ERK cell signaling pathway.Silencing GOLPH3 expression can reverse its resistance to some extent.