OCTN2 基因多态性影响结直肠癌奥沙利铂化疗敏感性  

OCTN2 gene polymorphisms affect the chemotherapeutic sensitivity of oxaliplatin in colorectal cancer

作　　者：鲁琼[1] 朱海宏 戚听听 李国华 滕欣琪 屈强[2] 屈健[1] LU Qiong;ZHU Haihong;QI Tingting;LI Guohua;TENG Xinqi;QU Qiang;QU Jian(Department of Pharmacy,the Second Xiangya Hospital,Central South University,Institute of Clinical Pharmacy,Central South University,Changsha 410011,Hunan,China;Department of Pharmacy,Xiangya Hospital,Central South University,Changsha 410078,Hunan,China)

机构地区：[1]中南大学湘雅二医院药剂科,中南大学临床药学研究所,湖南长沙410011 [2]中南大学湘雅医院药剂科,湖南长沙410078

出　　处：《中国临床药理学与治疗学》2021年第4期401-407,共7页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家自然科学基金(82073944,81603208);吴阶平医学基金(LCYX-Q001,320.6750.19090-11)。

摘　　要：目的:研究OCTN2基因多态性是否影响OCTN2的表达和功能,以及结直肠癌细胞系SW480对奥沙利铂敏感性。方法:构建OCTN2基因4种突变(F17L、E317K、S467C和P478L)转染细胞系。Real-time RT-PCR及Western blot检测OCTN2 mRNA和蛋白水平的表达。HPLC检测细胞内奥沙利铂含量。MTS法检测细胞活性。结果:所有突变型OCTN2 mRNA和蛋白表达水平与野生型表达无统计学差异。E317K和S467C细胞系对奥沙利铂摄取(V_(max)和K_(m))与野生型相比无差异(P>0.05),F17L的V_(max)下降到66.4%,而K_(m)值增加到120.3%,P478L的V_(max)增加到132.6%,K_(m)值降低到82.2%,与野生型相比有统计学差异(P<0.05)。所有过表达OCTN2细胞的IC 50值显著降低。与SW480-OCTN2转染细胞系比较,没有转染的SW480细胞和F17L细胞系的IC 50值分别升高175%和147%,而P478L细胞则下降到76.9%,有统计学差异(P<0.05)。E317K和S467C的IC 50值无明显改变(P>0.05)。结论:OCTN2转运体基因F17L和P478L错义突变可以改变OCTN2对奥沙利铂的摄取,从而影响OCTN2稳定表达细胞系对奥沙利铂的敏感性。AIM:To explore the effect of OCTN2 gene polymorphism on the expression and function of OCTN2,as well as the sensitivity of SW480 cells to oxaliplatin.METHODS:Four mutations of OCTN2(F17L,E317K,S467C and P478L)transfected cell lines were constructed.Real-time RT-PCR and Western blot were used to detect the levels of OCTN2 mRNA and protein.The content of oxaliplatin was detected by HPLC.MTS assay was used to detect cell viability.RESULTS:The expression level of all mutant OCTN2 mRNA and protein was not significantly different from that of wild-type OCTN2.Oxaliplatin uptake experiments showed that there was no significant difference in V_(max)and K_(m)values between E317K and S467C cells(P>0.05),the V_(max)of F17L decreased to 66.4%and K_(m)value increased to 120.3%;the V_(max)of P478L increased to 132.6%,and the value of K_(m)decreased to 82.2%,and there were significant differences(P<0.05).The cell activity test showed that the IC 50 value of all overexpressed OCTN2 cells decreased significantly.Compared with SW480-OCTN2 transfected cell lines,IC 50 values of SW480 cells without transfection and F17L cell lines were up to 175%and 147%,while P478L cells decreased to 76.9%,which had significant differences(P<0.05).The IC 50 value of E317K and S467C did not change significantly(P>0.05).CONCLUSION:The OCTN2 transporter gene F17L and P478L mutations can alter the uptake of oxaliplatin by OCTN2 transporter,thereby affecting the sensitivity of OCTN2-stably expression cell line to oxaliplatin.

关 键 词：结直肠癌 奥沙利铂 化疗敏感性 OCTN2转运体 

分 类 号：R735.3[医药卫生—肿瘤] R969[医药卫生—临床医学]