CAJ | 학술논문

目的检测放射诱导的肿瘤特异性嵌合启动子介导的辣根过氧化物酶/吲哚乙酸自杀基因系统对肺癌细胞A549、SPC-A1的生物效应.方法构建含有6个CArG元件的人端粒酶逆转录酶基因嵌合启动子调控的辣根过氧化物酶表达的质粒载体,及单个端粒酶逆转录酶启动子质粒和对照质粒,分别为pE6-hTERT-HRP、phTERT-HRP、pControl-HRP、pControl-luc.分别用细胞计数法、Annexin V-FITC染色检测此嵌合启动子质粒系统对肺癌细胞A549、SPC-A1和正常人胚肺细胞(hEL)的生长抑制及凋亡效应.利用成克隆分析法检测此系统对肺癌细胞A549、SPC-A1放射敏感性的影响.结果在6 Gy放射线诱导下质粒pE6-hTERT-HRP、phTERT-HRP、pControl-HRP、pControl-luc介导的自杀基因系统对A549细胞的平均生长抑制率分别为72.92%、40.60%、51.00%、25.19%(F=67.31,P<0.01),对SPC-A1细胞的平均抑制率分别为64.63%、30.02%、48.23%、23.16%(F=64.94,P<0.01),而对正常hEL细胞则分别为20.81%、18.05%、44.20%、18.32%(F=52.19,P<0.01).同样四种质粒对A549细胞的平均早期凋亡率分别为36.63%、22.30%、24.33%、12.53%(F=50.99,P<0.01),对SPC-A1细胞的平均早期凋亡率分别为33.73%、17.37%、22.43%、11.20%(F=20.76,P<0.01),而对正常hEL细胞则分别为13.53%、12.5%、21.93%、12.16%(F=15.08,P<0.01).同样四种质粒对A549细胞放射增敏比分别为3.45、2.29、3.05、1.21,对SPC-A1细胞的放射增敏比则分别为2.68、2.15、2.28、1.15.结论放射诱导的肿瘤特异性嵌合启动子介导的辣根过氧化物酶/吲哚乙酸自杀基因系统对肺癌细胞A549、SPC-A1具有特异杀伤作用,具有很好的应用前景.
목적 검측방사유도적종류특이성감합계동자개도적랄근과양화물매/신타을산자살기인계통대폐암세포A549、SPC-A1적생물효응.방법 구건함유6개CArG원건적인단립매역전록매기인감합계동자조공적랄근과양화물매표체적질립재체,급단개단립매역전록매계동자질립화대조질립,분별위pE6-hTERT-HRP、phTERT-HRP、pControl-HRP、pControl-luc.분별용세포계수법、Annexin V-FITC염색검측차감합계동자질립계통대폐암세포A549、SPC-A1화정상인배폐세포(hEL)적생장억제급조망효응.이용성극륭분석법검측차계통대폐암세포A549、SPC-A1방사민감성적영향.결과 재6 Gy방사선유도하질립pE6-hTERT-HRP、phTERT-HRP、pControl-HRP、pControl-luc개도적자살기인계통대A549세포적평균생장억제솔분별위72.92%、40.60%、51.00%、25.19%(F=67.31,P<0.01),대SPC-A1세포적평균억제솔분별위64.63%、30.02%、48.23%、23.16%(F=64.94,P<0.01),이대정상hEL세포칙분별위20.81%、18.05%、44.20%、18.32%(F=52.19,P<0.01).동양사충질립대A549세포적평균조기조망솔분별위36.63%、22.30%、24.33%、12.53%(F=50.99,P<0.01),대SPC-A1세포적평균조기조망솔분별위33.73%、17.37%、22.43%、11.20%(F=20.76,P<0.01),이대정상hEL세포칙분별위13.53%、12.5%、21.93%、12.16%(F=15.08,P<0.01).동양사충질립대A549세포방사증민비분별위3.45、2.29、3.05、1.21,대SPC-A1세포적방사증민비칙분별위2.68、2.15、2.28、1.15.결론 방사유도적종류특이성감합계동자개도적랄근과양화물매/신타을산자살기인계통대폐암세포A549、SPC-A1구유특이살상작용,구유흔호적응용전경.
Objective To detect specific cell killing effect of radiation combined with horseradish peroxidase (HRP)/indole-3-acetic (IAA) suicide gene therapy controlled by a novel radio-inducible and cancer-specific chimeric gene promoter in lung cancer. Methods We constructed a plasmid expressing HRP enzyme under the control of chimeric human telomerase reverse transcriptase (hTERT) promoter carrying 6 CArG elements, a plasmid expressing HRP enzyme under the control of hTERT promoter carrying single CArG element, and two control plasmids, which named pE6-hTERT-HRP, phTERT-HRP, pControl-HRP, and pControl-luc, respectively. After radiation, the proliferation inhibition and apoptosis induction effect of each type of plasmid in lung cancer cells (A549, SPC-A1) and normal lung cells (hEL) was detected by cell counting and Annexin V-FITC staining. The change of radiosensitivity of lung cancer cells with plasmid system was also detected by clonogenic assays. Results After a single dose radiation of 6 Gy,the average proliferation inhibition rates of pE6-hTERT-HRP, phTERT-HRP, pControl-HRP, and pControl-luc systems were 72. 92% ,40.60% , 51.00% and 25.19% (F= 67.31 , P< 0.01) in A549 cells ,64.63%,30.02%,48.23% and 23.16% (F=64.94, P< 0.01) in SPC-A1 cells, and 20.81%,18.05%, 44.20% and 18.32% (F=52. 19,P<0.01) in normal hEL cells, respectively. The average early apoptosis rates of these four plasmid systems were 36. 63%, 22. 30%, 24. 33% and 12. 53% (F =50. 99,P <0. 01) in A549 cells, 33.73%, 17. 37%, 22. 43% and 11.20% (F = 20. 76, P < 0. 01) in SPC-A1 cells, and 13.53 %, 12. 5%, 21.93% and 12. 16% (F = 15. 08, P < 0. 01) in normal hEL cells,respectively. The sensitizing enhancement ratios of the four plasmid systems were 3.45, 2. 29, 3.05 and 1.21 in A549 cells, while 2. 68, 2. 15, 3.05 and 1.21 in SPC-A1 cells, respectively. Conclusions The new suicide gene system controlled by chimeric promoter may provide a novel therapeutic modality for lung cancer.