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[目的]观察至真方含药血清对人大肠癌耐长春新碱细胞株HCT-8/VCR多药耐药的影响,并从NF-κB信号通路探讨其作用机制。[方法]用8%、16%、32%体积分数的至真方含药血清干预人大肠癌敏感细胞株HCT-8与耐长春新碱细胞株HCT-8/VCR,CCK8法检测细胞多药耐药性及存活率;ELISA法检测NF-κB活性;AnnexinV/PI流式细胞术检测细胞凋亡率;Western blot检测NF-κB/p65、IκB-α和Caspase-3蛋白的表达。[结果]体积分数>8%时,高、中、低剂量含药血清组对HCT-8/VCR细胞的抑制率与空白血清组比较差异均有统计学意义(P<0.01),且呈浓度递增趋势;8%、16%、32%体积分数的中剂量组至真方含药血清作用于HCT-8/VCR细胞24h后,NF-κB活性较HCT-8/VCR阴性对照组明显降低(P<0.01);至真方含药血清可促进HCT-8/VCR细胞凋亡,且在一定范围内细胞凋亡率呈浓度依赖性;经至真方含药血清处理后,HCT-8/VCR细胞NF-κB/p65蛋白表达较用药前明显下降(P<0.05),IκB-α蛋白表达明显上调(P<0.05),Caspase-3蛋白表达明显上调(P<0.05)。[结论]至真方含药血清可降低HCT-8/VCR细胞NF-κB的活性,一定程度上逆转了大肠癌细胞的多药耐药,其机制可能与抑制该通路分子开关IκB-α的磷酸化,下调NF-κB/p65蛋白的表达,从而上调通路下游的Caspase-3蛋白的表达,诱导HCT-8/VCR细胞凋亡有关。
[Objective] To observe the effect of ZJF serum on the multi-drug resistance of human colon carcinoma cell line VCR-HCT-8 / VCR and to explore its mechanism from NF-κB signaling pathway. [Method] HCT-8 and vincristine-resistant cell line HCT-8 / VCR were respectively treated with 8%, 16% and 32% Drug resistance and survival rate were measured. The activity of NF-κB was detected by ELISA. The apoptosis rate was detected by Annexin V / PI flow cytometry. The expressions of NF-κB / p65, IκB-α and Caspase-3 protein were detected by Western blot. [Result] When the volume fraction> 8%, the inhibitory rates of HCT-8 / VCR cells in high, medium and low dose serum groups were significantly different from those in blank serum group (P <0.01) (P <0.05). Compared with the HCT-8 / VCR negative control group, the activity of NF-κB in HCT-8 / VCR cells was significantly decreased after the medium dose of 8%, 16%, 32% P <0.01). To the serum containing HCT-8 / VCR, HCT-8 / VCR could induce apoptosis in a dose-dependent manner. The protein expression of NF-κB / p65 in VCR cells was significantly decreased (P <0.05), the expression of IκB-α was up-regulated (P <0.05) and the expression of Caspase-3 was up-regulated in VCR cells (P <0.05). [Conclusion] The serum containing Zhizhen Decoction can reduce the activity of NF-κB in HCT-8 / VCR cells and reverse the multidrug resistance of colorectal cancer cells to a certain extent. The mechanism may be related to the inhibition of IκB-α Phosphorylation and downregulate the expression of NF-κB / p65 protein, thereby upregulating the expression of Caspase-3 protein downstream of the pathway and inducing the apoptosis of HCT-8 / VCR cells.