过表达cMA3结构域增加黑色素瘤B16细胞对达卡巴嗪的药物敏感性

Drug sensitivity of melanoma B16 cells to dacarbazine was increased by overexpression of cMA3 domain

目的研究过表达c MA3结构域增强黑色素瘤B16细胞对达卡巴嗪(dacarbazine,DTIC)敏感性的影响及机制。方法利用脂质体转染使黑色素瘤B16细胞过表达c MA3结构域;使用实时PCR明确转染情况;分别使用CCK-8细胞增殖活力检测、流式细胞周期检测、流式细胞凋亡检测、Western-bloting凋亡蛋白检测转c MA3过表达质粒组与转空质粒组B16细胞在有无低剂量DTIC作用时的增殖活力、细胞周期、凋亡情况及凋亡蛋白的表达。结果转c MA3过表达质粒组的黑素瘤B16细胞的增殖活力可被低剂量(100μmol/L)DTIC抑制,而转空质粒组细胞的增殖活力不受影响。⑴B16细胞的细胞周期被低剂量DTIC阻滞于G2/M期;⑵B16细胞可被低剂量DTIC诱导凋亡;⑶B16细胞的凋亡相关蛋白表达可被低剂量DTIC影响。结论过表达c MA3结构域可以提高黑色素瘤B16细胞对DTIC的敏感性。

Objective To study the effect and mechanism of overexpression of c MA3 domain on the drug sensitivity of melanoma B16 cells to dacarbazine(DTIC).Methods Liposome transfection was used to overexpress the c MA3 domain in melanoma B16 cells;real-time PCR was used to confirm the status of overexpression;CCK-8 cell proliferation viability test,flow cytometry cell cycle test,flow cytometry apoptosis test and western-blotting apoptotic protein test were used to detect the proliferation viability,cell cycle,apoptotic status and apoptotic protein expression of B16 cells in the c MA3 overexpression group and the blank group with or without low-dose DTIC.Results In the c MA3 overexpression group,the proliferation viability of melanoma B16 cells could be inhibited by low-dose(100μmol/L)DTIC,but the the blank group was not affected.The cell cycle of B16 cells could be blocked in G2/M phase by low-dose DTIC in the c MA3 overexpression group.The apoptosis of B16 cells was induced by low-dose DTIC in the c MA3 overexpression group.The expression of apoptosis-related proteins in B16 cells could be affected by low-dose DTIC in the c MA3 overexpression group.Conclusion Overexpression of c MA3 domain could increase the sensitivity of melanoma B16 cells to DTIC.