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目的探讨肝癌射频消融(RFA)治疗后,外周血转化生长因子(TGF-β)对肿瘤组织中肝癌干细胞(LCSCs)发生和肝癌复发的影响。方法检测肝癌RFA治疗后的患者外周血TGF-β和肿瘤组织LCSCs的水平;体外实验研究亚致死温度对肝癌细胞系Hep1-6中肿瘤干细胞和TGF-β水平的影响。结果 RFA治疗后10例患者外周血TGF-β水平显著下降(Ⅰ型患者),另10例患者TGF-β水平显著增高(Ⅱ型患者)。Ⅰ、Ⅱ型患者肝癌复发时间分别为(2.45±1.46)和(11.90±3.72)个月,差异有统计学意义(P<0.001)。RFA治疗后,I型患者复发肝癌组织中CD13 mRNA的水平较治疗前明显上调,上调2倍以上;但RFA治疗对Ⅱ型患者CD13mRNA水平无明显影响。小鼠肝癌细胞系Hep1-6用45℃亚致死温度处理10 min后再继续培养24 h。45℃处理相比37℃处理,可显著上调CD13~+Hep1-6细胞水平(P<0.001),并且45℃处理导致Sox2表达增加和TGF-β表达下降。用中和抗体抑制TGF-β可显著增加CD13~+ Hep1-6细胞水平(P<0.001);而用Sox2 siRNA抑制Sox2表达则显著减少CD13~+ Hep1-6细胞水平(P<0.001)。结论 TGF-β可以通过抑制Sox2的表达来抑制LCSCs的产生,从而抑制肝癌复发。
Objective To investigate the effect of transforming growth factor-β (TGF-β) on the development of hepatocellular carcinoma (LCSCs) and recurrence of hepatocellular carcinoma after radiofrequency ablation (RFA) in liver cancer. Methods The levels of TGF-β and LCSCs in peripheral blood of patients with hepatocellular carcinoma after RFA were measured. The effects of sublethal temperature on the level of tumor stem cells and TGF-β in hepatocellular carcinoma cell line Hep1-6 were studied in vitro. Results The levels of TGF-β in peripheral blood were significantly decreased in 10 patients (type Ⅰ patients) after RFA treatment. TGF-β levels were significantly increased in the other 10 patients (type Ⅱ patients). The recurrence time of HCC in type Ⅰ and type Ⅱ patients was (2.45 ± 1.46) and (11.90 ± 3.72) months, respectively, with significant difference (P <0.001). After RFA treatment, the level of CD13 mRNA in type I patients with recurrent hepatocellular carcinoma was significantly up-regulated by more than 2 folds compared with that before treatment. However, the level of CD13 mRNA in type I patients was not significantly affected by RFA. The mouse hepatoma cell line Hep1-6 was treated with sub-lethal temperature of 45 ℃ for 10 min and then cultured for 24 h. Compared with 37 ℃ treatment, 45 ℃ treatment significantly increased the level of CD13 ~ + Hep1-6 cells (P <0.001), and 45 ℃ treatment resulted in the increase of Sox2 expression and the decrease of TGF-β expression. Inhibition of TGF-β by neutralizing antibody significantly increased the level of CD13 ~ + Hep1-6 cells (P <0.001). Sox2 siRNA inhibited the expression of Sox2 by significantly reducing the level of CD13 ~ + Hep1-6 cells (P <0.001). Conclusion TGF-β can inhibit the production of LCSCs by inhibiting the expression of Sox2, thereby inhibiting the recurrence of liver cancer.