受体依赖性TRAIL重组腺病毒对人肺癌细胞凋亡的诱导作用

目的：观察重组TRAIL腺病毒（Ad5TRAIL及Ad5F35TRAIL）对人非小细胞肺癌（nonsmallcell lung cancer，NSCLC）原代培养细胞和细胞株的凋亡诱导作用，探讨两种AdTRAIL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NCIH520及10例原代培养肺癌细胞中CAR和CD46的表达水平；分别以Ad5TRAIL及Ad5F35TRAIL重组腺病毒按MOI 10和50感染上述细胞，48 h后Annexin VFITC双标法流式细胞术检测细胞的早期凋亡。结果：A549、Z793、QG56和NCIH520 4株肺癌细胞中，CD46的表达均明显高于CAR表达。Z793、QG56细胞对Ad5TRAIL和Ad5F35TRAIL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±158）%、（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NCIH520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）；Ad5TRAIL和Ad5F35TRAIL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；Ad5TRAIL或Ad5F35TRAIL 感染后，5例的原代肺癌细胞检测到凋亡；与Ad5TRAIL相比，Ad5F35TRAIL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，Ad5F35TRAIL的作用强于Ad5TRAIL，更适合于非小细胞肺癌的基因治疗。

CAR or CD46 dependent TRAIL adenoviral vector induced apoptosis in lung cancer cells

Objective: To observe the effects of recombinant adenovirus TRAIL (AdS-TRAIL & Ad5F35-TRAIL) on apoptosis of non-small cell lung (NSCLC) cells, so as to assess the value of Ad-TRAIL in gene therapy of NSCLC. Meth-ods: CAR and CD46 expression levels in lung cancer cell lines (A549, Z793, QG56 and NCI-H520) 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 Ad5-TRAIL & Ad5F35-TRAIL adenoviral vectors at MOI 10 or 50, re-spectively; the percentage of apoptosis cells labeled by Annexin V-FITC 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 NCI-H520) and the primary lung cancer cells. Significant apoptosis was observed in Z793 and QG56 cells transfected with Ad5-TRAIL or Ad5F35-TRAIL at MOI 10, with the apoptosis rate being (1.76±2.10)% (Ad5-TRAIL), (15.96±2.89) % (Ad5F35-THAIL) and (6.05±1.58) % (Ad5-TRAIL), (10.11±1.26) % (Ad5F35-TRAIL), respectively, compared to no adenovirus-transfected cells (2.33±0.37] % and 5.95±1.89]%, respectively, P < 0.05). Less than 10% of apoptosis cells were detected in NCI-H520 cells transfected with Ad5- or Ad5F35-TRAIL at MOI 50 (12.89±3.2] % for AdS-TRAIL and 9.08±1.35]% for Ad5F35-TRAIL, respectively) compared to no adenovirus-transfected cells (7.04±2.17] %, P > 0.05). Moreover, apoptosis induced by Ad5- or Ad5F35-TRAIL 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 Ad5-TRAIL or Ad5F35-TRAIL vector. A higher percentage of apoptotic cells were found in Ad5F35-TRAIL group than those in Ad5-TRAIL and control groups. Conclusion: Ad5-TRAIL can induce apoptosis of NSCLC cells in vitro, and Ad5F35-TRAIL is more potent than Ad5-TRAIL, so Ad5F35-TRAIL is more suitable for gene therapy of NSCLC.