灵芝多糖联合5-FU对人结肠癌HCT-116细胞增殖及凋亡的影响

目的：探讨灵芝多糖(GLP)联合5-氟尿嘧啶(5-FU)对人结肠癌HCT-116细胞增殖抑制及凋亡的影响。方法：采用MTT法测定单独和联用GLP与5-FU对人结肠癌HCT-116细胞的协同抑制作用，应用联合指数法分析两药物之间相互作用，Hoechst33258荧光染色观察细胞的形态学改变，碘化吡啶(PI)标记流式细胞术(FCM)检测GLP联合5-FU对HCT-116细胞周期和凋亡抑制率的变化。结果：MTT法测定结果显示GLP和5-FU单独用药均能显著抑制HCT-116细胞增殖。GLP(＞659μg/mL)联用5-FU(＞1.6μg/mL)联用呈协同效应(CI＜1)，与5-FU单独用药组相比有极显著差异(P＜0.01)，并呈时间依赖性。给药48h后，Hoechst33258荧光染色可见典型的凋亡形态学特征。FCM检测凋亡显示两药联合作用24h后，5mg/mL GLP联合50μg/mL 5-FU的凋亡率为36.14%，高于5-FU(50μg/mL)单独用药组(26.07%)；1.25mg/mL GLP联合12.5μg/mL 5-FU、2.5mg/mL GLP联合25μg/mL 5-FU、5mg/mL GLP联合50μg/mL 5-FU作用48h后，凋亡率分别为20.95%、32.87%、30.01%，均高于5-FU(50μg/mL)单独用药组(14.51%)。联合用药组可使HCT-116细胞阻滞于S期。结论：较高质量浓度的GLP与5-FU联用具有协同抑制人结肠癌细胞株HCT-116细胞增殖，诱导细胞凋亡，阻滞细胞周期作用。

Effect of Combined Ganoderma lucidum Polysacchrides (GLP) and Fluorouracil on Proliferation and Apoptosis in Human Colon Carcinoma HCT-116 Cells

Objective: To explore the effect of Ganoderma lucidum polysacchrides (GLP) combined with fluorouracil treatment on the proliferation and apoptosis of human colon carcinoma HCT-116 cells in vitro. Methods: The inhibitory effects of both drugs on the proliferation of HCT-116 cells were measured by using MTT assay. The combinatorial index method was used to analyze their synergistic inhibitory effect. Observations of morphological changes were carried out with Hoechst33258 fluorescence staining. The impact of GLP on cell cycle and apoptosis rate of HCT-116 cells was evaluated by flow cytometry (FCM). Results: GLP and 5-FU treatment alone suppressed the proliferation of HCT-116 cells. GLP treatment (＞659 μg/mL) in combination with 5-FU (＞1.6 μg/mL) revealed an obvious synergistic effect on the proliferation of HCT-116 cells (CI＜1) which exhibited a significant difference when compared with 5-FU treatment alone (P＜0.01) in a time-dependent manner. After treatments for 48 h, the cells showed typical apoptosis characteristics. The apoptotic HCT-116 cells were 36.14% after GLP (5 mg/mL) combined with 5-FU (50μg/mL) treatment for 24 h, which was higher than that obtained from 5-FU treatment alone (26.07%). The apoptotic HCT-116 cells induced by 1.25 mg/mL GLP combined with 12.5μg/mL 5-FU, 2.5 mg/mL GLP combined with 25μg/mL 5-FU, and 5 mg/mL GLP combined with 50μg/mL 5-FU for 48 h were 20.95%, 32.87% and 30.01%, respectively, showing an increase compared with 5-FU (50 μg/mL) treatment alone (14.51%). The arrested cell cycle in combinatorial treatment group was observed at S phase. Conclusion: GLP at higher concentration combined with 5-FU can synergistically suppress the proliferation of human colon cancer HCT-116 cells, and enhanced apoptosis and cell cycle arrest in HCT-116 cells.