减毒鼠伤寒沙门菌Ⅲ型分泌系统介导Apoptin对肿瘤细胞糖酵解的影响

Effect of Apoptin Mediated by Attenuated Salmonella typhimurium Type Ⅲ Secretion System on Glycolysis in Tumor Cells

为进一步探讨重组减毒鼠伤寒沙门菌Ⅲ型分泌系统介导Apoptin对肿瘤细胞糖酵解的影响,将重组减毒鼠伤寒沙门菌∆crp∆asdSL1344(pYA3493-SopE-Apoptin)和互补菌株∆crp∆asdSL1344(pYA3493-SopE)分别于体外感染黑色素瘤B16F10细胞,于感染后12 h和24 h收集细胞,检测细胞内丙酮酸、乳酸含量以及丙酮酸激酶、己糖激酶的活性。结果显示,重组菌感染组能极显著下调丙酮酸含量(P<0.01);在感染24 h后,重组菌组乳酸含量极显著低于互补菌组和细胞对照组(P<0.01)。丙酮酸激酶和己糖激酶活性检测结果显示,重组菌组丙酮酸激酶活性和己糖激酶活性均显著低于互补菌组和细胞对照组(P<0.05)。以上结果表明,减毒鼠伤寒沙门菌Ⅲ型分泌系统介导Apoptin能抑制肿瘤细胞糖酵解的能力,通过下调丙酮酸和乳酸含量,降低丙酮酸激酶和己糖激酶活性,导致肿瘤细胞发生明显的代谢变化。

To further investigate the effect of Apoptin mediated by recombinant attenuated Salmonella typhimurium typeⅢsecretion system on glycolysis of tumor cells,B16F10 cells were infected with recombinant strain∆crp∆asdSL1344(pYA3493‐SopE‐Apoptin)or complementary strain∆crp∆asdSL1344(pYA3493‐SopE),and the cells were collected at 12 h and 24 h after infection to detect the levels of pyruvic acid,lactic acid,pyruvate kinase and hexokinase.The results showed that the intracellular pyruvate level of recombinant strain group was significantly reduced(P<0.01).At 24 h after infection,the lactic acid content of recombinant strain group was significantly lower than that of complementary strain group and cell control group(P<0.01).In addition,the pyruvate kinase activity and hexokinase activity of recombinant strain group were significantly lower than that of complementary strain group and cell control group(P<0.05).These results suggest that recombinant strain can inhibit the glycolysis ability of B16F10 cells by down‐regulating the pyruvate and lactic acid content and reducing the activities of pyruvate kinase and hexokinase,leading to significant metabolic changes in tumor cells.