基于CD14, IL-1β表达及SGC-7901细胞凋亡变化探讨清热化湿方防治胃癌的机制

Effect of Qingre Huashi Dectction on Regulating SGC-7901 Cell Apoptosis by Mediating CD-14 and IL-1β for Prevention and Treatment of Colon Cancer

摘要:

目的: 观察清热化湿方对人胃腺癌细胞7901增殖、凋亡及CD14基因、肿瘤坏死因子α(TNF-α)及白细胞介素1β(IL-1β)表达的影响。方法: 采用血清药理学方法,对照组予胎牛血清,治疗组分别予不同体积分数(5%,10%,15%,20%,30%)清热化湿方含药血清干预体外培养的胃腺癌细胞SGC-7901,应用四甲基偶氮唑盐微量酶反应比色法(MTT法)检测细胞增殖,流式细胞仪分析细胞凋亡率、细胞周期,免疫组化、Real-time PCR技术检测CD14,TNF-α,IL-1β蛋白及mRNA表达情况。结果: 与对照组比较,治疗组SGC-7901细胞存活率明显下降(P<0.05);与对照组比较,治疗组S期细胞减少、G₀/G₁期细胞增多,且与药物质量浓度有剂量依赖性(P<0.05)。治疗组可明显下调CD14,TNF-α,IL-1β蛋白及mRNA表达并促进细胞凋亡(P<0.05),以中剂量更为明显。结论: 清热化湿方可能通过调控CD14,TNF-α,IL-1β表达,介导SGC-7901细胞S期,诱导细胞凋亡,发挥防治胃癌的效应。

Abstract:

Objective: To investigate the intervention effect and potential mechanism of Qingre Huashi decoction(QHD) for preventing the onset of colon cancer by observing SGC-7901 cell proliferation and apoptosis along with the expression of CD14 gene, tumor necrosis factor alpha and interleukin 1 beta. Method: SGC-7901 cells were divided into control group, intervention group. In vitro, the control group was offered fetal calf serum. The intervention group was fed with serum containing QHD for 5%,10%,15%,20%,30%. The multiplication, proliferation and apoptosis of SGC-7901 cell were detected by MTT and flow cytometry respectively. The expression of CD14, TNF-α and IL-1β gene was assayed by immunohistochemistry and real-time PCR. Result: Compared with control group, the survival of SGC-7901 cell was decreased in QHD group(P<0.05). In QHD group, the cells in G₀/G₁ phase were increased but the S phase cell was on the decline. Those changes were correlated with the culturing content. There is a significant difference between QHD group and control group in those changes as mentioned above(P<0.05). Especially, the effect of middle concentration serum containing QHD on inducing cell apoptosis and down regulating the expression of CD14, TNF-α, IL-1β was obvious (P<0.05). Conclusion: QHD exerts a distinct effect on preventing the onset of colon cancer by mediating SGC-7901 cell S time, cell composition and regulating the expression of CD14, TNF-α and IL-1β and inducing cell apoptosis.