CLDN1影响放疗后肝癌侵袭转移潜能变化的机制研究

Study on the mechanism of CLDN1 affecting the invasion and metastasis potential of hepatocellular carcinoma after radiotherapy

[目的]分析放疗后肝癌侵袭转移能力及其相关分子标志物表达水平变化,探讨肝癌侵袭转移分子机理。[方法]随机将肝癌细胞分为放疗组(8 Gy射线照射)和未放疗组(未射线照射),检测HepG2和Huh7两组细胞的干性相关标记物(CD133、CD44)表达水平,并且通过蛋白印迹试验检测肝癌细胞放疗前后中CLDN1表达水平差异;通过Transwell小室试验检测放疗组及未放疗组肝癌细胞侵袭转移潜能,确定侵袭转移潜能改变趋向,并进一步探讨CLDN1在其中的作用。[结果]放疗后肝癌细胞发生侵袭转移数目较未放疗组明显增加(P<0.05),放疗后HepG2和Huh7肝癌细胞中CD133及CD44的表达量均显著高于未放疗组(P<0.05);CLDN1在放疗前后表达水平差异显著(P<0.05);而降低CLDN1的表达,肝癌细胞发生侵袭转移数量减少(P<0.05),并且放疗CLDN1低表达肝癌细胞,肝癌细胞表面干性相关标记物CD133、CD44表达上调表达不明显,与正常对照组(CLDN1未敲低组)比较差异有统计学意义(P<0.05)。[结论]放疗后肿瘤细胞干性增加,CLDN1可通过诱导肝癌细胞获得肿瘤干细胞特性,从而增强放疗后肝癌细胞侵袭转移能力。

[Objective]To analyze the changes of invasion and metastasis ability of hepatocellular carcinoma(HCC) and the expression level of related molecular markers after radiotherapy, and explore the molecular mechanism of invasion and metastasis of HCC. [Methods]Hepatocellular carcinoma cells were randomly divided into Radiotherapy group(8 Gy irradiated) and Non radiotherapy group(non-irradiated). The expression levels of dry stemness markers(CD133, CD44) in HepG2 and Huh7 cells were detected. The expression of CLDN1 in hepatocellular carcinoma cells before and after radiotherapy was detected by Western blotting. The invasion and metastasis potential of hepatocellular carcinoma cells in radiotherapy group and non-radiotherapy group were detected by Transwell lab experiment. Determine the change trend of invasion and metastasis potential, and further explore the role of CLDN1 in it. [Results]The number of invasion and metastasis of hepatocellular carcinoma cells after radiotherapy was significantly higher than that in non-radiotherapy group(P<0.05). The expression of CD133 and CD44 in HepG2 and Huh7 hepatocellular carcinoma cells after radiotherapy was significantly higher than that in non-irradiation group(P<0.05). The expression level of CLDN1 was significantly different before and after radiotherapy(P<0.05), but the number of invasion and metastasis of hepatocellular carcinoma cells was decreased by decreasing the expression of CLDN1(P<0.05). The expression of stemness related markers(CD133 and CD44)on the surface of hepatocellular carcinoma cells was not significantly up-regulated after radiotherapy, which was significantly different from that of normal control group(CLDN1 non-knockdown group)(P<0.05). [Conclusion]After radiotherapy, the stemness of tumor cells increased. CLDN1 can acquire the characteristics of cancer stem cells by inducing hepatocellular carcinoma cells,which can enhance the invasive and metastatic ability of hepatocellular carcinoma cells after radiotherapy.