中文摘要：

目的 观察荷载黑素瘤分化相关基因-7（mda-7）的复制缺陷性腺病毒Ad-mda-7联合氮烯咪胺（DTIC）对黑素瘤M14细胞的诱导凋亡作用.方法 将M14细胞分为4组进行干预,分别为PBS组、DTIC组、Ad-mda-7组、Ad-mda-7联合DTIC组.应用CCK-8法检测细胞抑制率;应用结晶紫法检测细胞毒性;应用Annexin Ⅴ法检测细胞凋亡率;应用Western blot法检测mda-7蛋白及bax、bcl-xL、Mcl-1、Caspase-9、Caspase-3凋亡相关蛋白表达.结果 CCK-8法显示10 MOI＋10 mg/L的Ad-mda-7联合DTIC干预M14细胞48 h后抑制率为（73.56±2.58）%,与对应剂量的Ad-mda-7（32.30±2.38）%、DTIC（15.75±3.21）%比较,差异有统计学意义（P＜0.05）;结晶紫法结果表明Ad-mda-7联合DTIC较Ad-mda-7、DTIC对M14细胞有显著的细胞毒性效应;Annexin Ⅴ法显示10MOI＋10 mg/L的Ad-mda-7联合DTIC干预M14细胞48 h后凋亡率为（71.47±3.57）%,与对应剂量的Ad-mda-7（40.11±3.61）%、DTIC（22.27±2.30）%比较,差异有统计学意义（P＜0.05）;Western blot结果表明Ad-mda-7联合DTIC作用的M14细胞能高效表达mda-7,同时bax的表达增加,Mcl-1、bcl-xL的表达降低,Caspase-9、Caspase-3被裂解活化.结论 荷载mda-7基因的复制缺陷性腺病毒Ad-mda-7联合化疗药物DTIC有明显的协同诱导黑素瘤细胞凋亡的作用.

英文摘要：

Objective To observe the effects of the combined use of replication-defective adenovirus carrying mda-7 （ Ad-mda-7） with Dacarbazine （DTIC） on the apoptosis of a melanoma cell line Ml4.Methods M14 cells were divided into 4 groups; PBS group, DTIC group, Ad-mda-7 group, Ad-mda-7 plus DTIC group. The inhibitory rate of cells was assessed by CCK-8 assay. Cell cytotoxic effect and apoptosis were determined by crystal violet staining method and Annexin V asssy, respectively. Furthermore, mda-7 proteins, bax, Mcl-1, bcl-xL, Caspase-9 and Caspase-3 apoptosis related proteins were determined by Western blotting. Results CCK-8 assay revealed the inhibitory rate after 48 h in the Ad-mda-7 （10 MOI） plus DTIC （10 mg/L） group （73.56 ±2.58）% was significantly higher than Ad-mda-7 group （33.30 ± 3.38 ） % and DTIC group （15.75 ± 3.21） % （ P 〈 0.05）. Crystal violet staining method showed Ad-mda-7 plus DTIC could induce more significant cytotoxiciy to M14 cells than Ad-mda-7 group or DTIC group. Annexin V assay indicated that the apoptosis rate after 48 h in Ad-mda-7 ＋ DTIC group （71.47 ± 3.57）% was significantly higher than in Ad-mda-7 group （40.11 ±3.61）% and DTIC group （23.27 ± 2.30）% （P〈0.05）. Western blotting analysis confirmed that M14 cells treated with Ad-mda-7 plus DTIC could express mda-7 with high efficiency, and simultaneously, the bax expression was increased, the expression of Mcl-1 and bcl-xL reduced and Caspase-9 and Caspase-3 were acitivated. Conclusion Ad-mda-7 in combination with DTIC exhibited a remarkably increased apoptosis-inducing effects in melanoma cells.