CAJ | 학술논문

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／L-OHP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。
목적：건립내오사리박（ L-OHP）적인결직장암내약세포주（ HT-29／L-OHP），검측상관내약기인LAPTM4β적표체，탐토기재L-OHP내약중적작용궤제。방법채용약물농도제도체증간헐유도법건립인결직장암내약주HT-29／L-OHP；응용LAPTM4β-35단백억제제작용우HT-29／L-OHP세포주후，류식세포기술검측세포조망，비교기여HT-29／L-OHP세포주대우L-OHP적내약정황；응용RT-PCR검측LAPTM4βmRNA적표체；응용Western blot측정LAPTM4β-35단백적표체。결과성공건립인결직장암내약세포주HT-29／L-OHP，류식세포기술검측결과표명응용LAPTM4β-35단백억제제작용우HT-29／L-OHP세포주후，기대우L-OHP적내약명현저우HT-29／L-OHP세포주。 HT-29／L-OHP세포주중LAPTM4βmRNA표체수평명현고우친본세포주HT-29，분별위5．613±0．139화1．105±0．187，（P＜0．05），LAPTM4β-35단백재HT-29／L-OHP세포화HT-29세포적상대표체량2．203±0．080화1．033±0．070，（P＜0．05）。결론 LAPTM4β재결직장암내약세포주중과표체，여결직장암오사리박내약밀절상관，가이작위결직장암내약적일개민감적생물학표지물。
Objective To establish L-OHP resistant human colorectal cancer cell line ( HT-29/L-OHP) , and detect the expression of related drug resistant gene LAPTM 4β, 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 LAPTM 4βmRNA;Western blot was used to detect the expression of LAPTM 4β-35 protein.Result L-OHP resistant human colorectal cancer cell line ( HT-29/L-OHP ) was established successfully .The flow cytometry result showed that LAPTM 4β-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 .