人宫颈癌紫杉醇耐药细胞株Hela/PTX的建立及其生物特性研究  

Establishment of Paclitaxel-resistance Cervical Carcinoma Cell Line Hela/PTX and Study on its Biological Characteristics

作　　者：李倩 王璇[1] 尹东锋[1] Li Qian;Wang Xuan;Yin Dongfeng(Pharmacy Department, General Hospital of Xinjiang Military Region, Urumqi 830000, Chin)

机构地区：[1]新疆军区总医院药剂科,乌鲁木齐830000

出　　处：《中国药师》2018年第5期787-791,共5页China Pharmacist

基　　金：国家自然科学基金项目(编号:81372782)

摘　　要：目的:建立人宫颈癌紫杉醇耐药细胞株Hela/PTX,并考察其生物学特性。方法:以人宫颈癌Hela细胞为亲本细胞,采用低浓度持续诱导法建立紫杉醇耐药细胞株Hela/PTX。在显微镜下观察细胞形态学的变化;采用CCK-8法检测耐药细胞株对不同化疗药物的耐药性;采用高效液相色谱法测定细胞中药物的蓄积;采用Western blot法和实时荧光定量PCR技术分别检测细胞中P糖蛋白(P-gp)、Survivin蛋白和mRNA表达。结果:成功建立Hela/PTX多药耐药细胞株,其对PTX的耐药数是58.40,且对多西他赛和氟尿嘧啶有交叉耐药,耐药指数分别为8.11,4.62。Hela/PTX细胞中P-gp和Survivin的mRNA表达水平分别是Hela细胞的(9.93±0.90)倍(P<0.05)、(3.02±0.57)倍(P<0.05)。在耐药细胞中P-gp和Survivin的蛋白表达水平明显高于亲本细胞。结论:成功构建了人宫颈癌耐药细胞株Hela/PTX,具有多药耐药的基本生物学特性,可做为紫杉醇耐药机制研究的体外模型及为宫颈癌多药耐药机制的研究提供实验基础。Objective： To establish a paclitaxel （PTX）-resistance cell line of human cervical carcinoma Hela/PTX and investigate its biological features. Methods： Using Hela as the negative cells, drug-resistant cervical cancer Hela/PTX cells were applied as the target cells. The morphology changes were observed under an optical microscope; the drug resistance of Hela/PTX to different chemotherapeutic drugs was detected by CCK-8 method. The accumulation of drugs in the cells was determined by HPLC. The expression of P-glycoprotein （P-gp）, Survivin protein and mRNA was detected by Western blot and RT-PCR. Results： Hela/PTX muhidrug resistant cell line was successfully established. The resistance index to PTX was 58.40 and the resistance index of cross-resistance to docetaxel and 5-fluorouracil was 8.11 and 4.62, respectively. The mRNA expression level of P-gp and Survivin protein in Hela/PTX cells respectively was （9.93 ± 0.90） times （P 〈 0.05） and （3.02 ±0.57） times （P 〈 0.05） of that in Hela cells. The expression of P-gp and Survivin protein in drug-resistant cells was significantly higher than that in parental cells. Conclusion： Human cervical cancer resistant cell line Hela/PTX with the basic biological characteristics of muhidrug resistance is successfully established, which can be used as an in vitro model for the studies on paclitaxel resistance mechanism and provide experimental basis for the studies of muhidrug resistance mechanism of cervical cancer.

关 键 词：紫杉醇 宫颈癌 多药耐药 HELA细胞株 

分 类 号：R965.1[医药卫生—药理学]