Tong Xuhui, Han Xi, Yu Binbin, Yu Meiling, Jiang Guojun, Ji Jie, Dong Shuying
Faculty of Pharmacy, Bengbu Medical College, Bengbu, Anhui 233030, P.R. China.
Oncol Rep. 2015 Jan;33(1):207-14. doi: 10.3892/or.2014.3571. Epub 2014 Oct 29.
Platinum agents are widely used in the chemotherapy of testicular cancer. However, adverse reactions and resistance to such agents have limited their application in antineoplastic treatment. The aim of the present study was to determine the role of gap junction intercellular communication (GJIC) composed of Cx43 on oxaliplatin‑induced survival/apoptosis in mouse leydig normal and cancer cells using MTT, Annexin V/PI double staining assays and western blot analysis. The results showed that GJIC exerted opposite effects on the mouse leydig cancer (I-10) and normal (TM3) cell apoptosis induced by oxaliplatin. In leydig cancer cells, survival of cells exposed to oxaliplatin was substantially reduced when gap junctions formed as compared to no gap junctions. Pharmacological inhibition of gap junctions by oleamide and 18-α-glycyrrhetinic acid resulted in enhanced survival/decreased apoptosis while enhancement of gap junctions by retinoic acid led to decreased survival/increased apoptosis. These effects occurred only in high‑density cultures (gap junction formed), while the pharmacological modulations had no effects when there was no opportunity for gap junction formation. Notably, GJIC played an opposite (protective) role in normal leydig cells survival/apoptosis following exposure to oxaliplatin. Furthermore, this converse oxaliplatin‑inducing apoptosis exerted through the functional gap junction was correlated with the mitochondrial pathway‑related protein Bcl-2/Bax and caspase‑3/9. These results suggested that in testicular leydig normal/cancer cells, GJIC plays an opposite role in oxaliplatin‑induced apoptosis via the mitochondrial pathway.
铂类药物广泛应用于睾丸癌的化疗。然而，这些药物的不良反应和耐药性限制了它们在抗肿瘤治疗中的应用。本研究的目的是使用MTT、Annexin V/PI双染法和蛋白质印迹分析，确定由Cx43组成的间隙连接细胞间通讯（GJIC）在奥沙利铂诱导的小鼠睾丸间质正常细胞和癌细胞存活/凋亡中的作用。结果表明，GJIC对奥沙利铂诱导的小鼠睾丸间质癌细胞（I-10）和正常细胞（TM3）凋亡具有相反的作用。在睾丸间质癌细胞中，与未形成间隙连接相比，形成间隙连接时暴露于奥沙利铂的细胞存活率显著降低。油酰胺和18-α-甘草次酸对间隙连接的药理学抑制导致存活率提高/凋亡减少，而视黄酸增强间隙连接则导致存活率降低/凋亡增加。这些效应仅发生在高密度培养（形成间隙连接）时，而当没有机会形成间隙连接时，药理学调节没有作用。值得注意的是，GJIC在正常睾丸间质细胞暴露于奥沙利铂后的存活/凋亡中起相反（保护）作用。此外，通过功能性间隙连接产生的这种相反的奥沙利铂诱导凋亡与线粒体途径相关蛋白Bcl-2/Bax和caspase-3/9相关。这些结果表明，在睾丸间质正常/癌细胞中，GJIC通过线粒体途径在奥沙利铂诱导的凋亡中起相反作用。
