左金丸对胃癌耐药细胞SGC-7901/DDP增殖和糖酵解的抑制作用

Zuojin Pill inhibits proliferation and glycolysis in DDP-resistant gastric cancer cell SGC-7901/DDP

目的:研究左金丸对胃癌耐药细胞SGC-7901/DDP增殖和糖酵解的影响。方法:取人胃癌细胞株SGC-7901,体外诱导顺铂(DDP)耐药细胞株SGC-7901/DDP。将耐药细胞分为空白对照组和左金丸低、中、高浓度(25、50、100μg/ml)组,各组予以相应干预。药物干预12 h、24 h、48 h后,CCK-8分别检测各组细胞存活率;药物干预24 h后,试剂盒检测各组细胞葡萄糖摄取率及细胞上清中乳酸含量,同时应用Western blot检测各组细胞中糖酵解途径关键分子C-myc、缺氧诱导因子-1α(HIF-1α)、葡萄糖转运蛋白1 (GLUT1)、乳酸脱氢酶A(LDHA)、己糖激酶Ⅱ(HKⅡ)的蛋白表达。结果:(1)与对照组比较,左金丸各浓度组在各检测时间点的细胞存活率均显著降低(P<0.01),且呈时间、浓度依赖性,提示左金丸可显著抑制SGC-7901/DDP细胞增殖。(2)药物干预24 h后,与对照组比较,左金丸各浓度组细胞的葡萄糖摄取率及乳酸含量显著降低(P<0.01),且呈浓度依赖性,提示左金丸可显著抑制SGC-7901/DDP糖酵解。(3)不同浓度左金丸干预24 h后,SGC-7901/DDP细胞中C-myc、HIF-1α、GLUT1、LDHA、HKⅡ的蛋白表达显著下调(P<0.01),且呈浓度依赖性。结论:左金丸可显著抑制胃癌耐药细胞SGC-7901/DDP的增殖,且这一过程可能与抑制糖酵解相关。

Objective: To study the effects of Zuojin Pill on the proliferation and glycolysis of cisplatin( DDP)-resistant gastric cancer cell line SGC-7901/DDP. Methods: Human gastric cancer cell line SGC-7901 was taken,and the DDP-resistant cell line SGC-7901/DDP was induced in vitro. The resistant cells were divided into the blank control group and Zuojin Pill groups with low-,medium-and high-concentration( 25,50,100 μg/ml). Each group was treated with the corresponding intervention. After treatment for 12,24 and 48 hours,CCK8 assay was used to evaluate the cell survival rate. After treatment for 24 hours,the glucose uptake and the lactic acid content in cell supernatant were detected by kit. Meanwhile,the expressions of proliferation and glycolysis related proteins,including C-myc,hypoxia-inducible factor-1α( HIF-1α),glucose transporter 1( GLUT1),lactate dehydrogenase A( LDHA),and hexokinaseⅡ( HKⅡ) were detected by Western blot. Results:(1)Compared with the control group,the survival rates of Zuojin Pill groups with different concentrations were decreased significantly at each timepoint( P<0.01),in a time-and concentration-dependent manner,which suggested that Zuojin Pill could significantly inhibit the cell proliferation of SGC-7901/DDP.(2)After treatment for 24 hours,compared with the control group,the levels of glucose uptake and lactic acid content were significantly reduced in Zuojin Pill groups with different concentrations( P<0.01),in a concentration-dependent manner,which suggested that Zuojin Pill could significantly inhibit the glycolysis of SGC-7901/DDP.(3)After treatment for 24 hours,the expressions of C-myc,HIF-1α,GLUT1,LDHA and HK Ⅱ were significantly down-regulated in Zuojin Pill groups with different concentrations( P<0.01),in a concentration-dependent manner. Conclusion: Zuojin Pill can significantly inhibit the proliferation of DDP-resistant gastric cancer cell SGC-7901/DDP,which may be related to the inhibition on glycolysis.