魔芋葡甘露聚糖逆转大肠癌5-氟尿嘧啶耐药的作用及其机制研究

Study on Effect and Its Mechanism of Konjac Glucomannan on the Reversal of 5-Fluorotracicl Resistance in Colorectal Cancer

目的探究魔芋葡甘露聚糖(konjac glucomannan,KGM)体内外逆转大肠癌5-氟尿嘧啶(5-fluorouracil, 5-FU)耐药作用及其相关机制。方法构建HCT-8/5-FU耐药细胞株,采用MTT法验证HCT-8/5-FU细胞的耐药性以及KGM和5-FU对HCT-8/5-FU细胞的增殖抑制作用;采用CompuSyn软件分析KGM和5-FU联合用药的联合指数,评价联合效应;流式细胞术检测两药对HCT-8/5-FU细胞凋亡的影响;Westernblotting检测PI3K/Akt信号通路蛋白表达情况;构建裸鼠移植瘤模型,检测KGM和5-FU体内抑瘤效果。结果 HCT-8/5-FU细胞株对5-FU的敏感性显著降低;KGM和5-FU单用均能够抑制HCT-8/5-FU细胞的增殖,两者联用能够更加有效地抑制HCT-8/5-FU细胞增殖;CompuSyn软件分析结果表明KGM和5-FU具有协同作用;KGM和5-FU联用能够显著增加HCT-8/5-FU细胞凋亡率,并抑制PI3K/Akt信号通路活性(P<0.05);KGM和5-FU联用的抑瘤率显著高于5-FU单独使用(P<0.05)。结论 KGM能够逆转大肠癌5-FU耐药,其作用机制跟PI3K/Akt信号通路有关。

OBJECTIVE To investigate the reversal effect of 5-fluorouracil(5-FU) resistance of colorectal cancer by konjac glucomannan(KGM) in vivo and in vitro and its related mechanism. METHODS HCT-8/5-FU resistant cell lines were constructed, and MTT assay was used to verify the resistance of HCT-8/5-FU cells and the inhibitory effect of KGM and 5-FU on proliferation of HCT-8/5-FU cells. The combined index of KGM and 5-FU was analyzed by CompuSyn software to evaluate the combined effect. The effects of the two drugs on apoptosis were detected by flow cytometry. Expression of PI3 K/Akt signaling pathway was detected by Western blotting. The transplanted tumor model of nude mice was constructed to detect the tumor inhibition effect of KGM and 5-FU in vivo. RESULTS The sensitivity of HCT-8/5-FU cell line to 5-FU was significantly decreased. Both KGM and 5-FU alone could inhibit the proliferation of HCT-8/5-FU cells, while the combination of the two could inhibit the proliferation of HCT-8/5-FU cells more effectively. CompuSyn software analysis results showed that KGM and 5-FU had a synergistic effect. KGM combined with 5-FU significantly increased the apoptosis rate of HCT-8/5-FU and inhibited the activity of PI3 K/Akt signaling pathway(P<0.05). The tumor inhibition rate of KGM combined with 5-FU was significantly higher than that of 5-FU alone(P<0.05). CONCLUSION KGM can reverse 5-FU resistance in colorectal cancer, and its mechanism is related to the PI3 K/Akt signaling pathway.