MDA-7/IL-24对食管癌细胞的增殖和存活影响

Adenovirus Vector Expressing MDA-7/IL-24 Induces Growth Arrests and Apoptosis in Human Esophageal Cancer Cell Lines

目的：研究黑色瘤分化相关基因‐7（MDA‐7）／白细胞介素‐24（IL‐24）对食管癌细胞 Eca‐109与 TE‐1的增殖抑制作用和杀伤作用。方法应用RT‐PCR技术检测MDA‐7／IL‐24受体复合物在Eca‐109与 TE‐1细胞的表达，构建MDA‐7／IL‐24基因的重组腺病毒载体，将携带人MDA‐7／IL‐24基因的腺病毒Ad‐mda‐7分别感染食管癌细胞Eca‐109与TE‐1，通过Western‐blot检测MDA‐7基因的表达，通过CCK‐8检测该基因对食管癌细胞的增殖抑制作用，碘化丙啶（PI）染色后流式细胞仪检测MDA‐7对食管癌细胞周期的影响，Hoechst／PI双染色后流式细胞仪观察MDA‐7对食管癌细胞的诱导凋亡作用。结果 Eca‐109与 TE‐1均有表达MDA‐7／IL‐24受体复合物IL‐22R1／IL‐20R2；成功构建包装重组腺病毒，并经 Western‐blot验证其介导了外源基因 MDA‐7／IL‐24在Eca‐109与TE‐1细胞中的高效表达；CCK‐8实验结果表明，MDA‐7／IL‐24能显著抑制Eca‐109与 TE‐1细胞的增殖，且随着感染时间的延长及感染剂量的增加而增强抑制作用；细胞周期结果提示，MDA‐7／IL‐24使大量Eca‐109与TE‐1细胞被阻滞在G1～G0期；Hoechst／PI双染色结果提示，MDA‐7／IL‐24能明显促进Eca‐109与TE‐1细胞的凋亡。结论重组腺病毒能介导MDA‐7／IL‐24基因通过G1期阻滞和诱导凋亡，从而抑制食管癌细胞的增殖。

Objective To study the effects of melanoma differentiation associated gene‐7/interleu‐kin 24 （MDA‐7/IL‐24） induction of grow th arrests and apoptosis on human esophageal cancer cell lines , Eca‐109 and TE‐1 . Methods The mRNA expressions of MDA‐7/IL‐24 receptor complexes in Eca‐109 and TE‐1 were confirmed using RT‐PCR . The MDA‐7/IL‐24 gene was transfected into Eca‐109 and TE‐1 with adenovirus vector . Protein expression was confirmed using Western‐blot . CCK‐8 assay and flow cytometry were used to analysis tumor cell proliferation and cell cycle separately . Flow cytometry assay after Hoechst and PI staining was performed to analysis the apoptosis . Results MDA‐7/IL‐24 receptor complexes IL‐22R1/IL‐20R2 was expressed in both Eca‐109 and TE‐1 . The protein product of MDA‐7/IL‐24 was detected in Eca‐109 and TE‐1 . MDA‐7/IL‐24 effectively inhibited the growth and proliferation of Eca‐109 and TE‐1 with the extension of time or the increase of MOI . Furthermore ,MDA‐7/IL‐24 induced G1 -G0 growth arrest and apoptosis in Eca‐109 and TE‐1 . Conclusion MDA‐7/IL‐24 inhibited the proliferation by inducing G1 -G0 growth arrest and apoptosis in Eca‐109 and TE‐1 . This suggestive MDA‐7/IL‐24 can be a perfect gene for gene therapy in esophageal carcinoma .