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目的:探讨鼻咽癌CD133+干细胞中ABCG2表达情况及CD133+干细胞对肿瘤多药耐药的作用。方法:免疫磁珠分选、纯化鼻咽癌CD133+肿瘤干细胞,采用无血清培养、CCK-8法、平板克隆及裸鼠体内成瘤实验鉴定此肿瘤干细胞生物学特性;采用逆转录聚合酶链式反应(RT-PCR)和免疫组化检测分选前后的ABCG2/BCRP变化;采用CCK-8法比较分选前后鼻咽癌细胞的耐药情况。结果:成功分选并鉴定CD133+鼻咽癌干细胞;RT-PCR结果显示,耐药基因ABCG2在CD133+干细胞中表达灰度值为2.27±0.09,明显高于CD133-(0.69±0.01)和未分选细胞(1.00±0.00),F=886.99,P<0.01;CCK8结果显示,顺铂、紫杉醇和依托泊苷对CD133+鼻咽癌细胞的耐药明显增加,耐药指数分别为23.874、5.520和5.670,P均<0.01,而氯丙嗪的耐药指数为1.082,差异无统计学意义,P=0.488。结论:CD133+干细胞参与鼻咽癌多药耐药,靶向ABCG2联合氯丙嗪有可能成为克服肿瘤多药耐药新途径。
Objective: To investigate the expression of ABCG2 in nasopharyngeal carcinoma CD133 + stem cells and the effect of CD133 + stem cells on multidrug resistance of tumor. METHODS: Nasopharyngeal carcinoma CD133 + tumor stem cells were sorted by immunomagnetic beads. The biological characteristics of the tumor stem cells were identified by serum-free culture, CCK-8 assay, plate clone and nude mice in vivo. Reverse transcription polymerase chain reaction The changes of ABCG2 / BCRP before and after the sorting were detected by RT-PCR and immunohistochemistry. The drug resistance of nasopharyngeal carcinoma cells before and after the sorting was compared by CCK-8 method. Results: The CD133 + NPC stem cells were successfully sorted and identified. RT-PCR results showed that the gray value of ABCG2 in CD133 + stem cells was 2.27 ± 0.09, which was significantly higher than that of CD133- (0.69 ± 0.01) and non-sorted (1.00 ± 0.00), F = 886.99, P <0.01. The results of CCK8 showed that the resistance of CD133 + NPC cells to cisplatin, paclitaxel and etoposide significantly increased, and the resistance index was 23.874, 5.520 and 5.670, P <0.01, while the resistance index of chlorpromazine was 1.082, the difference was not statistically significant, P = 0.488. Conclusion: CD133 + stem cells are involved in multidrug resistance of nasopharyngeal carcinoma. Targeting ABCG2 combined with chlorpromazine may become a new way to overcome the multidrug resistance of tumor.