[关键词]
[摘要]
目的:观察重组TRAIL腺病毒(Ad5TRAIL及Ad5F35TRAIL)对人非小细胞肺癌(nonsmallcell lung cancer,NSCLC)原代培养细胞和细胞株的凋亡诱导作用,探讨两种AdTRAIL用于肺癌基因治疗的价值。方法:采用流式细胞术检测人肺癌细胞系A549、Z793、QG56和NCIH520及10例原代培养肺癌细胞中CAR和CD46的表达水平;分别以Ad5TRAIL及Ad5F35TRAIL重组腺病毒按MOI 10和50感染上述细胞,48 h后Annexin VFITC双标法流式细胞术检测细胞的早期凋亡。结果:A549、Z793、QG56和NCIH520 4株肺癌细胞中,CD46的表达均明显高于CAR表达。Z793、QG56细胞对Ad5TRAIL和Ad5F35TRAIL的作用较敏感, MOI=10感染后凋亡率分别为(11.76±2.10)%、(15.96±2.89)%和(6.05±158)%、(10.11±1.26)%,显著高于对照组\[(2.33±0.37)%和(5.95±1.89)%,P<0.05\];MOI=50感染时NCIH520细胞凋亡率分别为(12.89±3.2)%和(9.08±1.35)%,与对照组(7.04±2.17)%相比差异无统计学意义(P>0.05);Ad5TRAIL和Ad5F35TRAIL均不能诱导A549细胞凋亡。10例原代肺癌细胞CD46表达也明显较CAR高;Ad5TRAIL或Ad5F35TRAIL 感染后,5例的原代肺癌细胞检测到凋亡;与Ad5TRAIL相比,Ad5F35TRAIL诱导的凋亡率更高。结论:两种TRAIL重组腺病毒对非小细胞肺癌细胞均有凋亡诱导作用,Ad5F35TRAIL的作用强于Ad5TRAIL,更适合于非小细胞肺癌的基因治疗。
[Key word]
[Abstract]
Objective:To observe the effects of recombinant adenovirus TRAIL (Ad5TRAIL & Ad5F35TRAIL) on apoptosis of nonsmall cell lung (NSCLC) cells, so as to assess the value of AdTRAIL in gene therapy of NSCLC. :Methods: :CAR and CD46 expression levels in lung cancer cell lines (A549, Z793, QG56 and NCIH520) and the primary lung cancer cells from samples of 10 NSCLC patients were assayed by flow cytometry analysis. The lung cancer cell lines and primary lung cancer cells were infected with Ad5TRAIL & Ad5F35TRAIL adenoviral vectors at MOI 10 or 50, respectively; the percentage of apoptosis cells labeled by Annexin VFITC in different cells were measured by flow cytometry 48 h after transfection. :Results:: The expression of CD46 were higher than that of CAR in all the lung cancer lines (A549, Z793, QG56 and NCIH520) and the primary lung cancer cells. Significant apoptosis was observed in Z793 and QG56 cells transfected with Ad5TRAIL or Ad5F35TRAIL at MOI 10, with the apoptosis rate being (1.76±2.10)%(Ad5TRAIL), (15.96±2.89)%(Ad5F35TRAIL)and (6.05±1.58)%(Ad5TRAIL),(10.11±1.26)%(Ad5F35TRAIL), respectively, compared to no adenovirustransfected cells (\[2.33±0.37\]% and \[5.95±1.89\]%, respectively, P<0.05). Less than 10% of apoptosis cells were detected in NCIH520 cells transfected with Ad5 or Ad5F35TRAIL at MOI 50 (\[12.89±3.2\]% for Ad5TRAIL and \[9.08±1.35\]% for Ad5F35TRAIL, respectively)compared to no adenovirustransfected cells (\[7.04±2.17\]%, P>0.05). Moreover, apoptosis induced by Ad5 or Ad5F35TRAIL transfection in A549 cells was not detected both at MOI 10 and 50. About half of the primary lung cancer cells from 10 patients induced apoptosis after transfected with Ad5TRAIL or Ad5F35TRAIL vector. A higher percentage of apoptotic cells were found in Ad5F35TRAIL group than those in Ad5TRAIL and control groups. :Conclusion: :Ad5TRAIL can induce apoptosis of NSCLC cells in vitro, and Ad5F35TRAIL is more potent than Ad5TRAIL, so Ad5F35TRAIL is more suitable for gene therapy of NSCLC.
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[基金项目]
江苏省社会发展基金资助项目(No.BS2007025)