LAPTM4β在人结直肠癌耐药细胞株中的表达及意义  

Expression and significance of LAPTM4β in L-OHP resistant human colorectal cancer cell line

作　　者：刘家麒[1] 宁丽[2] 李云龙[3] 游柳平 王春敬[1] 黄跃南[1] 

机构地区：[1]哈尔滨医科大学附属第二医院普外科,150081 [2]哈尔滨医科大学附属第二医院妇产科,150081 [3]哈尔滨医科大学附属第二医院重症监护病房,150081

出　　处：《中华结直肠疾病电子杂志》2015年第5期50-54,共5页Chinese Journal of Colorectal Diseases(Electronic Edition)

基　　金：黑龙江省教育厅科学技术项目(12541303)

摘　　要：目的建立耐奥沙利铂(L-OHP)的人结直肠癌耐药细胞株(HT-29/L-OHP),检测相关耐药基因LAPTM4β的表达,探讨其在L-OHP耐药中的作用机制。方法采用药物浓度梯度递增间歇诱导法建立人结直肠癌耐药株HT-29/L-OHP;应用LAPTM4β-35蛋白抑制剂作用于HT-29/L-OHP细胞株后,流式细胞技术检测细胞凋亡,比较其与HT-29/L-OHP细胞株对于L-OHP的耐药情况;应用RT-PCR检测LAPTM4βmRNA的表达;应用Western blot测定LAPTM4β-35蛋白的表达。结果成功建立人结直肠癌耐药细胞株HT-29/L-OHP,流式细胞技术检测结果表明应用LAPTM4β-35蛋白抑制剂作用于HT-29/L-OHP细胞株后,其对于L-OHP的耐药明显低于HT-29/L-OHP细胞株。HT-29/LOHP细胞株中LAPTM4βmRNA表达水平明显高于亲本细胞株HT-29,分别为5.613±0.139和1.105±0.187,(P<0.05),LAPTM4β-35蛋白在HT-29/L-OHP细胞和HT-29细胞的相对表达量2.203±0.080和1.033±0.070,(P<0.05)。结论 LAPTM4β在结直肠癌耐药细胞株中过表达,与结直肠癌奥沙利铂耐药密切相关,可以作为结直肠癌耐药的一个敏感的生物学标志物。Objective To establish L-OHP resistant human colorectal cancer cell line( HT-29 /LOHP),and detect the expression of related drug resistant gene LAPTM4β,and probe its role in the mechanism of L-OHP resistant.Method Drug concentration gradient increasing intermittent induction method was used to establish L-OHP resistant human colorectal cancer cell line( HT-29 / L-OHP);LAPTM4β-35 protein inhibitor was used in HT-29 / L-OHP cell line,flow cytometry was adopted to detect cell apoptosis,and to compare L-OHP resistant of it with other HT-29 / L-OHP cell line;RT-PCR was adopted to detect the expression of LAPTM4βmRNA;Western blot was used to detect the expression of LAPTM4β-35 protein.Result L-OHP resistant human colorectal cancer cell line( HT-29 / L-OHP) was established successfully.The flow cytometry result showed that LAPTM4β-35 protein inhibitor use in HT-29 / L-OHP cell line lead to significantly lower L-OHP resistance than that of HT-29 / L-OHP cell line.The expression of LAPTM4βmRNA of HT-29 / L-OHP was significantly higher than parental cell plant HT-29,5.613 + 0.139 and 1.105 + 0.187,respectively( P < 0.05).The relative expression level of LAPTM4-35 in HT-29 / L-OHP cell line and HT-29 cell line was 2.203 + 0.080 and 1.033 + 0.070,( P < 0.05).Conclusion LAPTM4βis overexpressed in HT-29 / L-OHP cell line,and it is closely related with L-OHP resistant of colorectal cancer,it could be used as a sensitive biomarker for drug resistance of colorectal cancer.

关 键 词：结直肠肿瘤 抗药性 肿瘤 奥沙利铂 LAPTM4β 

分 类 号：R735.34[医药卫生—肿瘤]