槐耳菌质对人胆管癌细胞侵袭与迁移的影响及其作用机制

Effect of Huai'erjunzhi on invasion and migration of human cholangiocarcinoma cells and its action mechanism

背景与目的:胆管癌作为罕见的恶性肿瘤,难以诊断,往往在晚期才被发现,只能选择姑息疗法,但是常规的化疗药物对胆管癌治疗效果较差,急需寻找新的治疗药物。本研究探讨槐耳菌质对人胆管癌细胞恶性生物学行为的影响,及其与TGF-β/Smad通路的关系,旨在为槐耳菌质治疗胆管癌提供理论依据。方法:用不同浓度槐耳菌质孵育人正常肝细胞L-02和人胆管癌细胞HuCCT1不同时间后,检测细胞增殖情况,并计算半数抑制浓度(IC_(50));将HuCCT1细胞分为阴性对照组(无干预)、阳性对照组(15 mg/L顺铂)和不同浓度槐耳菌质干预组(根据浓度与时间为预实验结果选择1/5 IC_(50)、2/5 IC_(50)和IC_(50)),用划痕试验与Transwell实验检测细胞迁移与侵袭能力,同时用Western blot检测HuCCT1细胞中TGF-β/Smad通路相关蛋白的表达。结果:只有较高浓度的槐耳菌质(>312.5 mg/L)对L-02细胞的增殖有明显抑制作用;当浓度超过5 mg/L时,槐耳菌质能明显抑制HuCCT1细胞增殖,且呈浓度依赖性(均P<0.05),IC_(50)分别为24 h:138.52 mg/L、48 h:99.41 mg/L、72 h:113.52 mg/L。与阴性对照组比较,阳性对照组与三种浓度(20、40、100 mg/L)的槐耳菌质干预组HuCCT1细胞迁移距离、侵袭细胞数减少,TGF-β1、Smad2、Smad3、Smad4、N-cadherin、Snail和Slug表达降低,E-cadherin表达增加(均P<0.05);与阳性对照组比较,三个槐耳菌质干预组的上述变化更弱,但呈明显的浓度依赖性(均P<0.05)。结论:槐耳菌质可以抑制HuCCT1细胞恶性生物学行为,其作用机制可能与抑制TGF-β/Smad通路活性有关。

Background and Aims:Cholangiocarcinoma,a rare malignant tumor,is difficult to diagnose and often detected at an advanced stage,limiting treatment options to palliative care.Conventional chemotherapy drugs have poor efficacy against cholangiocarcinoma,making the search for new treatments critical.This study was conducted to investigate the effects of Huai'erjunzhi on the malignant biological behavior of human cholangiocarcinoma cells and its relationship with the TGF-β/Smad pathway,aiming to provide a theoretical basis for the use of Huai'erjunzhi in cholangiocarcinoma treatment.Methods:Human normal liver cells(L-02)and human cholangiocarcinoma cells(HuCCT1)were incubated with different concentrations of Huai'erjunzhi for various durations.Cell proliferation was assessed,and the half-maximal inhibitory concentration(IC_(50))was calculated.HuCCT1 cells were divided into a negative control group(no intervention),a positive control group(15 mg/L cisplatin),and different Huai'erjunzhi intervention groups(1/5 IC_(50),2/5 IC_(50),and IC_(50) based on preliminary experimental results).Scratch and Transwell assays were used to measure cell migration and invasion,while Western blot was employed to detect the expression of proteins related to the TGF-β/Smad pathway in HuCCT1 cells.Results:Only high concentrations of Huai'erjunzhi(>312.5 mg/L)significantly inhibited the proliferation of L-02 cells.Huai'erjunzhi significantly inhibited the proliferation of HuCCT1 cells at concentrations above 5 mg/L in a concentration-dependent manner(all P<0.05),with IC_(50) values of 138.52 mg/L at 24 h,99.41 mg/L at 48 h,and 113.52 mg/L at 72 h.Compared with the negative control group,the positive control group and the three Huai'erjunzhi intervention groups(20,40,and 100 mg/L)exhibited reduced migration distance,decreased invasive cell numbers,lower expression of TGF-β1,Smad2,Smad3,Smad4,N-cadherin,Snail,and Slug,and increased expression of E-cadherin(all P<0.05).Compared with the positive control group,these changes in the Huai'erjunzhi groups were less pronounced but showed a clear concentration-dependent relationship(all P<0.05).Conclusion:Huai'erjunzhi can potentially inhibit the malignant biological behavior of HuCCT1 cells by inhibiting the TGF-β/Smad pathway.