Objective: To investigate the effect of transcatheter arterial chemoembolization(TACE)combined with thymosin alpha1(Tα1)on the autophagy of immune cells from advanced hepatocellular carcinoma. Methods: A total of 30 patients with advanced liver cancer enrolled in Lishui Central Hospital from September 2015 to June 2016 were collected in this study. The average age of patients was 16-75(56±12) years. All patients were treated with TACE after enrolled in hospital in a week. Patients were divided into TACE group and TACE+ Tα1 treatment group(15 cases in each group). Patients in TACE group received a conventional treatment, without any immunotherapy, while the TACE+ Tα1 treatment group accepted TACE following a subcutaneously injection of 1.6 mg Tα1 twice a week for 4 weeks. Flow cytometry was used to detect the T cell subsets in two groups both before and after TACE treatment for 1, 4 weeks and at 3 months follow-up. Peripheral blood mononuclear cells (PBMC) were isolated by density gradient centrifugation. The expression of Beclin-1, LC3 protein and mRNA were detected by Western blot (WB) and PCR respectively. Results: There was no statistical difference of the percentage of CD3(+) , CD4(+) , CD8(+) T cell subsets and Beclin-1, LC3 protein and mRNA expression between the two groups before TACE treatment (P>0.05). The percentage of CD3(+) , CD4(+) , CD8(+) T cell subsets in TACE+ Tα1 group at 1 week post-TACE treatment (58.45%±16.34%, 38.33%±15.16%, 27.31%±12.54%), at 4 weeks post-TACE treatment (62.38%±18.62%, 43.19%±13.86%, 29.54%±10.33%) and 3 months follow-up (64.15%±13.76%, 41.28%±14.65%, 29.38%±15.65%) were statistically higher than those in TACE group at 1 week post-TACE treatment (53.71%±11.17%, 32.12%±10.53%, 24.45%±13.72%) at 4 weeks post-TACE treatment (52.12%±14.26%, 31.16%±15.43%, 23.39%±15.33%) and 3 months follow-up (54.28%±13.15%, 32.17%±14.98%, 24.34%±14.12%) (P<0.05). The Beclin-1, LC3 protein and mRNA expression in TACE+ Tα1 group at 1 week post-TACE treatment (protein: 0.57±0.08, 2.26±0.36, mRNA: 0.62±0.11, 2.69±0.27), at 4 weeks post-TACE treatment (protein: 0.66±0.09, 3.11±0.45, mRNA: 0.78±0.13, 3.43±0.61) were higher than those in TACE group at 1 week post-TACE treatment (protein: 0.45±0.16, 1.43±0.30, mRNA: 0.52±0.15, 1.15±0.37), at 4 weeks post-TACE treatment (protein: 0.51±0.13, 1.81±0.35, mRNA: 0.56±0.10, 1.98±0.41) ( P<0.05). But there was no statistically significant difference in the expression of Beclin-1 and LC3 in two groups at 3 months follow-up (P>0.05). Conclusions: TACE combined with Tα1 significantly increase the level of autophagy in the immune cells of patients with advanced primary hepatocellular carcinoma.
目的： 探讨肝动脉化疗栓塞术(TACE)联合胸腺肽α1(Tα1)治疗对中晚期肝癌患者免疫细胞自噬水平的改变。 方法： 收集2015年9月至2016年6月在丽水市中心医院住院的中晚期肝癌患者30例，年龄16～75(56±12)岁，所有研究对象均行TACE治疗，随机分为TACE组和TACE＋胸腺肽组(每组15例)，TACE组术后予常规处理；TACE＋胸腺肽组在TACE术后当天即刻给予1.6 mg Tα1皮下注射，每周2次，给药间隔3～4 d，共4周为1个疗程。用流式细胞仪检测两组患者TACE术前、术后1、4周及随访3个月时T细胞亚群百分比。采用密度－梯度离心法分离患者外周血单个核细胞(PBMC)，并用蛋白免疫印迹技术(WB)和PCR技术检测PBMC的Beclin－1、LC3蛋白和mRNA水平。 结果： 两组TACE术前CD3(＋)、CD4(＋)、CD8(＋)T细胞亚群百分比，Beclin－1、LC3蛋白和mRNA差异均无统计意义学(均P>0.05)。TACE＋胸腺肽组在术后1周时(58.45%±16.34%、38.33%±15.16%、27.31%±12.54%)、术后4周时(62.38%±18.62%、43.19%±13.86%、29.54%±10.33%)和随访3个月时(64.15%±13.76%、41.28%±14.65%、29.38%±15.65%)的CD3(＋)、CD4(＋)、CD8(＋) T细胞亚群百分比均明显增高于TACE组在术后1周时(53.71%±11.17%、32.12%±10.53%、24.45%±13.72%)、术后4周时(52.12%±14.26%、31.16%±15.43%、23.39%±15.33%)和术后3个月时(54.28%±13.15%、32.17%±14.98%、24.34%±14.12%)的百分比(均P<0.05) 。TACE＋胸腺肽组在术后1周时(蛋白表达：0.57±0.08、2.26±0.36，mRNA表达：0.62±0.11、2.69±0.27)和术后4周时(蛋白表达：0.66±0.09、3.11±0.45，mRNA表达：0.78±0.13、3.43±0.61)Beclin－1、LC3蛋白和mRNA表达均明显高于TACE组在术后1周时(蛋白表达：0.45±0.16、1.43±0.30，mRNA表达：0.52±0.15、1.15±0.37)和术后4周时(蛋白表达：0.51±0.13、1.81±0.35，mRNA表达：0.56±0.10、1.98±0.41)的表达(均P<0.05)；但随访3个月时表达差异无统计学意义(P>0.05)。 结论： TACE联合Tα1可明显增加中晚期原发性肝癌患者免疫细胞自噬水平。.
Keywords: Immunity, cellular; Liver neoplasms; Microtubule-associated protein light chain 3; Thymosin.