目的： 构建表达Bcl-XL小发夹RNA的腺病毒载体（Ad/Bcl-XL shRNA）并探讨其抗肿瘤作用。 方法： 构建、纯化重组腺病毒Ad/Bcl-XL shRNA。通过Western blotting、MTT分析验证它对Bcl-XL的下调及其杀伤肿瘤细胞的作用，并检测其处理后细胞凋亡信号的活化情况；在裸鼠皮下荷瘤模型中验证其体内抗肿瘤作用。 结果： 成功构建和纯化了Ad/Bcl-XL shRNA，它能显著下调结肠癌DLD1细胞Bcl-XL蛋白的表达；与Ad/GFP、PBS组相比，Ad/Bcl-XL shRNA组明显抑制人结肠癌细胞DLD1的生长\[1 000 MOI时(60.6±4.8)% vs (99.0±2.6)%、100%; 2 000 MOI时, (37.3±6.9)% vs (99.0±2.1)%、100%, P<0.01\]，但对正常人成纤维细胞无明显抑制作用(P>0.05)；Ad/Bcl-XL shRNA组能有效诱导结肠癌细胞中凋亡信号casepase-9、casepase-3、PARP的活化。在裸鼠荷瘤模型中，与Ad/GFP、PBS组相比，Ad/Bcl-XL shRNA组显著抑制DLD1来源皮下肿瘤的生长\[第29天时，（250.1±185.7) vs (880.0±286.1)、(911.0±389.1) mm 3；P<0.01\]。 结论： Ad/Bcl-XL shRNA能显著抑制结肠癌细胞在体内外的生长，其在结肠癌治疗中具有潜在的应用价值。

Objective : To construct the adenovector expressing small hairpin RNA targeting Bcl-XL (Ad/Bcl-XL shRNA ), and evaluate its anti-tumor effect. Methods: Firstly, Ad/Bcl-XL shRNA was constructed and purified. Then the protein level of Bcl-XL and survival of colon cancer cells after the treatment of Ad/Bcl-XL shRNA were determined by Western blotting and MTT assay, respectively. Furthermore, the activation of apoptotic signaling was also detected by Western blotting assay. Finally, the anticancer effect of Ad/Bcl-XL shRNA in vivo was confirmed in the subcutaneous tumor model derived from DLD1 cells in nude mice. Results: Ad/Bcl-XL shRNA was constructed and purified successfully. It obviously down-regulated the Bcl-XL protein and significantly inhibited the growth of DLD1 cells (1 000 MOI and 2 000 MOI Ad/Bcl-XL shRNA group was (MOI=1 000: \[60.6±4.8\]% vs \[37.3±6.9\]%， 100%; MOI=2 000: \[99.0±26\]% vs \[ 99.0± 2.1\]%, 100% P<0.01), but had no obvious toxicity on normal human fibroblasts. Western blotting results demonstrated that the apoptotic signal molecules including caspase-9， caspase-3, and PARP were obviously activated after the treatment with Ad/Bcl-XL shRNA. In vivo, it also dramatically suppressed the growth of subcutaneous tumors derived from DLD1 cells in nude mice (eg.29th day Ad/Bcl-XL shRNA group was \[250.1±185.7\] vs Ad/GFP \[880.0±286.1\]，PBS \[ 9110± 389.1\] mm 3，P<0.01). Conclusion: Ad/Bcl-XL shRNA can down-regulate the expression of Bcl-XL and inhibit the growth of colon cancer cells in vivo and in vitro, suggesting that it may be a new strategy to treat the colon carcinoma.

国家自然科学基金资助项目（No. 30700970）