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目的研究门冬酰胺酶(asparaginase,AN)对小细胞肺癌H1688和H446细胞的抗癌作用,探讨自噬在AN抗癌过程中的功能。方法采用MTT法及台盼蓝染色法检测AN单独或联用自噬抑制剂氯喹(chloroquine,CQ)对H1688和H446细胞的生长抑制作用;免疫荧光法观察自噬标记物LC3表达,示踪自噬体形成;Western blot检测自噬相关蛋白LC3及Akt/m TOR信号通路的表达。结果 AN呈浓度依赖性抑制H1688和H446细胞增殖并促进其死亡(P<0.05);AN处理组可以显著增加H1688和H446细胞自噬小体数量并诱导LC3-Ⅱ表达;自噬抑制剂CQ提高AN对H1688和H446细胞杀伤作用(P<0.05);AN作用H1688细胞后p-Akt、p-m TOR、p-70S6K蛋白表达降低。结论 AN对小细胞肺癌H1688和H446细胞具有抑制作用并诱导细胞保护性自噬,阻断自噬可以增强AN对小细胞肺癌H1688和H446细胞的抗癌疗效。
Objective To study the anti-cancer effect of asparaginase (AN) on small cell lung cancer H1688 and H446 cells and to explore the function of autophagy in the anti-cancer process of AN. Methods MTT assay and Trypan blue staining were used to detect the inhibitory effect of chloroquine (CQ) on the growth of H1688 and H446 cells. The expression of LC3 was detected by immunofluorescence, The formation of phage was detected by Western blot. The expressions of LC3 and Akt / m TOR signaling pathways were detected by Western blot. Results AN could inhibit the proliferation and promote the death of H1688 and H446 cells in a concentration-dependent manner (P <0.05). The AN group could significantly increase the number of autophagosomes and induce the expression of LC3-Ⅱ in H1688 and H446 cells; AN on H1688 and H446 cells (P <0.05); The expression of p-Akt, pm TOR, p-70S6K in H1688 cells treated with AN was decreased. Conclusions AN has inhibitory effect on small cell lung cancer H1688 and H446 cells and induces protective autophagy. Blocking autophagy can enhance antitumor effect of AN on small cell lung cancer H1688 and H446 cells.