中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2015年
24期
1951-1956
,共6页
申聪香%刘艳慧%文忠%杨柯柯%李冠雪%张沈华%张鑫雨
申聰香%劉豔慧%文忠%楊柯柯%李冠雪%張瀋華%張鑫雨
신총향%류염혜%문충%양가가%리관설%장침화%장흠우
鼻咽肿瘤%化疗%PinX1基因%端粒酶
鼻嚥腫瘤%化療%PinX1基因%耑粒酶
비인종류%화료%PinX1기인%단립매
Nasopharyngeal neoplasms%Chemotherapy%PinX1 gene%Telomerase
目的 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.方法 采用慢病毒构建的pCDH-CMV-PinX1-copGFP质粒载体转染鼻咽癌细胞,构建含PinX1基因载体的鼻咽癌Lenti-PinX1-5-8F细胞,同时构建Lenti-Ctrl-5-8F细胞(将不含PinX1基因的空载体转染5-8F细胞获得)及5-8F细胞作对照.将上述鼻咽癌细胞依实验分为4组:实验组为Lenti-PinX1-5-8F细胞+顺铂组(含PinX1基因的Lenti-PinX1-5-8F细胞中加入顺铂药物),对照组为Lenti-PinX1-5-8F细胞组(含PinX1基因)、顺铂药物组(5-8F细胞中加入顺铂)及5-8F细胞组.分别采用荧光定量PCR、流式细胞术、四甲基偶氮唑蓝(MTT)比色法、小室技术、Western印迹法及药物敏感试验检测PinX1基因表达、端粒酶活性、鼻咽癌增殖抑制、与顺铂抗癌的协同作用及肺耐药相关蛋白(LRP)及B细胞淋巴瘤基因-2(Bcl-2)基因的表达,观察PinX1基因在鼻咽癌细胞中对顺铂化疗敏感性的影响及其机制.结果 Lenti-PinX1-5-8F细胞中端粒酶活性相对量均显著低于Lenti-Ctrl-5-8F细胞、5-8F细胞(0.146±0.004比0.967 ±0.016、1.000±0.034,均P＜0.01).16 μg/ml的顺铂与PinX1基因联合能显著增强端粒酶活性下调后对鼻咽癌细胞的抑制作用.Lenti-PinX1-5-8F细胞+顺铂组的细胞增殖指数均显著低于Lenti-PinX1-5-8F细胞组、顺铂药物组、5-8F细胞组[(14.39±3.66)％比(32.97±3.00)％、(31.18±4.24)％、(47.19±4.19)％,均P＜0.01].Lenti-PinX1-5-8F细胞中LRP、Bcl-2蛋白相对表达量分别为0.64±0.14、0.57 ±0.12,均显著低于Lenti-Ctrl-5-8F细胞的0.84±0.19、0.81±0.16和5-8F细胞的0.83±0.35、0.78±0.27(均P＜0.01).结论 PinX1基因能增强顺铂对鼻咽癌细胞的化疗敏感性,其作用有可能是通过下调鼻咽癌细胞端粒酶活性、抑制Bcl-2和LRP基因而实现的.
目的 探討PinX1基因對鼻嚥癌細胞順鉑化療敏感性的影響及其機製.方法 採用慢病毒構建的pCDH-CMV-PinX1-copGFP質粒載體轉染鼻嚥癌細胞,構建含PinX1基因載體的鼻嚥癌Lenti-PinX1-5-8F細胞,同時構建Lenti-Ctrl-5-8F細胞(將不含PinX1基因的空載體轉染5-8F細胞穫得)及5-8F細胞作對照.將上述鼻嚥癌細胞依實驗分為4組:實驗組為Lenti-PinX1-5-8F細胞+順鉑組(含PinX1基因的Lenti-PinX1-5-8F細胞中加入順鉑藥物),對照組為Lenti-PinX1-5-8F細胞組(含PinX1基因)、順鉑藥物組(5-8F細胞中加入順鉑)及5-8F細胞組.分彆採用熒光定量PCR、流式細胞術、四甲基偶氮唑藍(MTT)比色法、小室技術、Western印跡法及藥物敏感試驗檢測PinX1基因錶達、耑粒酶活性、鼻嚥癌增殖抑製、與順鉑抗癌的協同作用及肺耐藥相關蛋白(LRP)及B細胞淋巴瘤基因-2(Bcl-2)基因的錶達,觀察PinX1基因在鼻嚥癌細胞中對順鉑化療敏感性的影響及其機製.結果 Lenti-PinX1-5-8F細胞中耑粒酶活性相對量均顯著低于Lenti-Ctrl-5-8F細胞、5-8F細胞(0.146±0.004比0.967 ±0.016、1.000±0.034,均P＜0.01).16 μg/ml的順鉑與PinX1基因聯閤能顯著增彊耑粒酶活性下調後對鼻嚥癌細胞的抑製作用.Lenti-PinX1-5-8F細胞+順鉑組的細胞增殖指數均顯著低于Lenti-PinX1-5-8F細胞組、順鉑藥物組、5-8F細胞組[(14.39±3.66)％比(32.97±3.00)％、(31.18±4.24)％、(47.19±4.19)％,均P＜0.01].Lenti-PinX1-5-8F細胞中LRP、Bcl-2蛋白相對錶達量分彆為0.64±0.14、0.57 ±0.12,均顯著低于Lenti-Ctrl-5-8F細胞的0.84±0.19、0.81±0.16和5-8F細胞的0.83±0.35、0.78±0.27(均P＜0.01).結論 PinX1基因能增彊順鉑對鼻嚥癌細胞的化療敏感性,其作用有可能是通過下調鼻嚥癌細胞耑粒酶活性、抑製Bcl-2和LRP基因而實現的.
목적 탐토PinX1기인대비인암세포순박화료민감성적영향급기궤제.방법 채용만병독구건적pCDH-CMV-PinX1-copGFP질립재체전염비인암세포,구건함PinX1기인재체적비인암Lenti-PinX1-5-8F세포,동시구건Lenti-Ctrl-5-8F세포(장불함PinX1기인적공재체전염5-8F세포획득)급5-8F세포작대조.장상술비인암세포의실험분위4조:실험조위Lenti-PinX1-5-8F세포+순박조(함PinX1기인적Lenti-PinX1-5-8F세포중가입순박약물),대조조위Lenti-PinX1-5-8F세포조(함PinX1기인)、순박약물조(5-8F세포중가입순박)급5-8F세포조.분별채용형광정량PCR、류식세포술、사갑기우담서람(MTT)비색법、소실기술、Western인적법급약물민감시험검측PinX1기인표체、단립매활성、비인암증식억제、여순박항암적협동작용급폐내약상관단백(LRP)급B세포림파류기인-2(Bcl-2)기인적표체,관찰PinX1기인재비인암세포중대순박화료민감성적영향급기궤제.결과 Lenti-PinX1-5-8F세포중단립매활성상대량균현저저우Lenti-Ctrl-5-8F세포、5-8F세포(0.146±0.004비0.967 ±0.016、1.000±0.034,균P＜0.01).16 μg/ml적순박여PinX1기인연합능현저증강단립매활성하조후대비인암세포적억제작용.Lenti-PinX1-5-8F세포+순박조적세포증식지수균현저저우Lenti-PinX1-5-8F세포조、순박약물조、5-8F세포조[(14.39±3.66)％비(32.97±3.00)％、(31.18±4.24)％、(47.19±4.19)％,균P＜0.01].Lenti-PinX1-5-8F세포중LRP、Bcl-2단백상대표체량분별위0.64±0.14、0.57 ±0.12,균현저저우Lenti-Ctrl-5-8F세포적0.84±0.19、0.81±0.16화5-8F세포적0.83±0.35、0.78±0.27(균P＜0.01).결론 PinX1기인능증강순박대비인암세포적화료민감성,기작용유가능시통과하조비인암세포단립매활성、억제Bcl-2화LRP기인이실현적.
Objective To explore the influence and mechanism of PinX1 gene on the chemotherapy sensitivity of nasopharyngeal carcinoma cells in response to Cisplatin.Methods Transfeeted nasopharyngeal carcinoma 5-8F cell lines with pCDH-CMV-PinX1-copGFP vector constructed by lentivirus to generate LentiPinX1-5-8F 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.Results 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-PinXl-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 (LentiCtrl-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).Conclusions 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.
