Shen Congxiang, Liu Yanhui, Wen Zhong, Yang Keke, Li Guanxue, Zhang Shenhua, Zhang Xinyu
Department of Otolaryngology, Pearl River Hospital, Southern Medical University, Guangzhou 510282, China.
Department of Otolaryngology, Pearl River Hospital, Southern Medical University, Guangzhou 510282, China; Email:
Zhonghua Yi Xue Za Zhi. 2015 Jun 23;95(24):1951-6.
To explore the influence and mechanism of PinX1 gene on the chemotherapy sensitivity of nasopharyngeal carcinoma cells in response to Cisplatin.
Transfected nasopharyngeal carcinoma 5-8F cell lines with pCDH-CMV-PinX1-copGFP vector constructed by lentivirus to generate Lenti-PinX1-5-8F cells containing PinX1 gene, using Lenti-Ctrl-5-8F cell (blank vector without PinX1 gene was used to transfect 5-8F cell lines) and 5-8F cell as controls. Expression of PinX1 gene, telomerase activity, the inhibition of cancer cells proliferation, combined anticancer effect with Cisplatin and the expression of lung resistance protein (LRP) and Bcl-2 were detected with fluorescent quantitation polymerase chain reaction (PCR), flow cytometry, thiazolyl blue (MTT) method, areole test, Western blot and drug sensitivity test, respectively, in four groups (Lenti-PinX1-5-8F cell + Cisplatin, Lenti-PinX1-5-8F cell, Cisplatin and 5-8F cell) so as to explore the influence and mechanism of PinX1 gene on the chemotherapy sensitivity of nasopharyngeal carcinoma cells in response to Cisplatin.
The telomerase activity in Lenti-PinX1-5-8F cell (0.146 ± 0.004) was lower than those in the other two control cells (Lenti-Ctrl-5-8F cell: 0.967 ± 0.016, 5-8F cell: 1.000 ± 0.034, both P < 0.01). The cancer cell biological activity could be intensively inhibited by 16 µg/ml Cisplatin after lower level telomerase activity induced by PinX1 gene. Proliferation index (PI) (%) in Lenti-PinX1-5-8F cell + Cisplatin (14.39 ± 3.66) was also less than the other groups (Lenti-PinX1-5-8F cell, Cisplatin and 5-8F cell groups, 32.97 ± 3.00, 31.18 ± 4.24 and 47.19 ± 4.19, all P < 0.01). And same time, the expressions of LRP (0.64 ± 0.14) and Bcl-2 (0.57 ± 0.12) protein in Lenti-PinX1-5-8F cells were obviously reduced than those in other two group cells (Lenti-Ctrl-5-8F cell: 0.84 ± 0.19 and 0.81 ± 0.16; 5-8F cell: 0.83 ± 0.35 and 0.78 ± 0.27; all P < 0.01).
PinX1 gene can enhance the chemotherapy sensitivity of nasopharyngeal carcinoma cells in response to Cisplatin, which may be mediated by the down-regulation of telomerase activity and the inhibition of LRP and Bcl-2 gene in nasopharyngeal carcinoma cells.
探讨PinX1基因对鼻咽癌细胞顺铂化疗敏感性的影响及其机制。
用慢病毒构建的pCDH-CMV-PinX1-copGFP载体转染鼻咽癌5-8F细胞系，构建含PinX1基因的Lenti-PinX1-5-8F细胞，以Lenti-Ctrl-5-8F细胞（用不含PinX1基因的空载体转染5-8F细胞系）和5-8F细胞作为对照。分别采用荧光定量聚合酶链反应（PCR）、流式细胞术、噻唑蓝（MTT）法、光晕试验、蛋白质免疫印迹法及药敏试验，检测四组细胞（Lenti-PinX1-5-8F细胞+顺铂、Lenti-PinX1-5-8F细胞、顺铂及5-8F细胞）中PinX1基因表达、端粒酶活性、癌细胞增殖抑制率、与顺铂联合抗癌效应以及肺耐药蛋白（LRP）和Bcl-2的表达，以探讨PinX1基因对鼻咽癌细胞顺铂化疗敏感性的影响及其机制。
Lenti-PinX1-5-8F细胞的端粒酶活性（0.146±0.004）低于其他两个对照细胞（Lenti-Ctrl-5-8F细胞：0.967±0.016，5-8F细胞：1.000±0.034，均P<0.01）。PinX1基因诱导端粒酶活性降低后，16μg/ml顺铂可显著抑制癌细胞生物学活性。Lenti-PinX1-5-8F细胞+顺铂组的增殖指数（PI）（%）（14.39±3.66）也低于其他组（Lenti-PinX1-5-8F细胞、顺铂及5-8F细胞组，分别为32.97±3.00、31.18±4.24及47.19±4.19，均P<0.01）。同时，Lenti-PinX1-5-8F细胞中LRP蛋白（0.64±0.14）和Bcl-2蛋白（0.57±0.12）的表达明显低于其他两组细胞（Lenti-Ctrl-5-8F细胞：0.84±0.19和0.81±0.16；5-8F细胞：0.83±0.35和0.78±0.27；均P<0.01）。
PinX1基因可增强鼻咽癌细胞对顺铂的化疗敏感性，其机制可能与下调鼻咽癌细胞端粒酶活性及抑制LRP和Bcl-2基因有关。
