人结肠癌细胞HCT-8/V及HCT-8中葡萄糖神经酰胺合成酶的表达

Expression of glucosylceramide synthase in human colon cancer cells HCT-8/V and HCT-8

目的:观察人结肠癌HCT-8细胞和其耐药细胞株HCT-8/V中葡萄糖神经酰胺合成酶(GCS)的表达及意义。方法:体外培养HCT-8细胞和HCT-8/V,采用CCK8法检测2种细胞对长春新碱(VCR)、5-氟尿嘧啶(5-Fu)及顺铂(DDP)的耐药性,计算IC50。采用免疫细胞化学法和RT-PCR法检测2种细胞中GCS蛋白及mRNA的表达。结果:HCT-8/VCR细胞对VCR、5-Fu和DDP的IC50均高于HCT-8细胞[(25.42±0.03)vs(0.63±0.02),t=1190.873,P<0.001;(18.25±0.02)vs(0.87±0.01),t=1346.249,P<0.001;(37.12±0.01)vs(1.59±0.03),t=1946.058,P<0.001)];HCT-8/V细胞中GCS蛋白阳性区平均积分光密度值(119.40±1.59)高于HCT-8细胞的(90.59±1.12),HCT-8/V细胞中GCSmRNA的表达(0.79±0.06)高于HCT-8细胞的(0.33±0.04),差异均有统计学意义(t=25.658,2.776,P均<0.05)。结论:GCS的高表达可能参与了人结肠癌多药耐药的形成。

Aim：To explore the expression and significance of glucosylceramide synthase（GCS）in human colon cancer cell line HCT-8 and its drug-resistant subline cell HCT-8/V.Methods：HCT-8 and HCT-8/V were cultured in vitro.CCK-8（Cell Counting kit-8）was used to detect the drug resistance of the two cells to VCR,5-Fu and DDP,and calculate IC50.The expressions of GCS protein and mRNA in the two cells were assayed by immunocytochemistry and RT-PCR,