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目的:探讨人反义端粒酶RNA(human telomerase RNA,hTR)对肝癌细胞系BEL-7402黏附和侵袭的影响及其机制。方法:利用前期实验成功转染端粒酶正、反义RNA基因的肝癌细胞,分为正义转染组(BEL-7402-hTR-EcoRⅠ细胞)、反义转染组(BEL-7402-hTR-BamHⅠ细胞),空白对照组(BEL-7402细胞)。黏附实验和侵袭实验检测反义hTR转染后BEL-7402细胞的黏附和侵袭能力,免疫细胞化学检测整合素β1(integrinβ1,INTβ1)、上皮钙黏蛋白(epithelial cadherin,E-cad)的表达水平,明胶酶谱法检测基质金属蛋白酶(matrix metalloproteinase,MMP)-2、-9的活性变化。结果:与空白对照组和正义hTR转染组相比,反义hTR转染组BEL-7402-hTR-BamHⅠ细胞的黏附、侵袭能力明显减低(0.204±0.029vs0.505±0.037、0.465±0.029,34.80±3.19vs71.47±5.15、69.87±3.11,均P<0.05),INTβ1表达降低(0.153±0.016vs0.385±0.008、0.375±0.014,P<0.05),E-cad表达增强(0.209±0.020vs0.124±0.018、0.134±0.016,P<0.05),MMP-2和MMP-9的活性受到明显抑制(2 054.22±138.52vs3 105.56±329.60、2 923.22±269.08,846.33±104.66vs1 538.89±122.85、1 453.33±126.35,均P<0.05)。结论:人反义端粒酶RNA能明显减低肝癌BEL-7402细胞的黏附、侵袭能力,其机制可能与上调E-cad的表达、下调INTβ1的表达,并抑制MMP-2和MMP-9的活性有关。
Objective: To investigate the effect of human telomerase RNA (hTR) on the adhesion and invasion of hepatocellular carcinoma cell line BEL-7402 and its mechanism. Methods: The hepatoma cells transfected with positive and negative telomerase telomerase genes were successfully transfected with BEL-7402-hTR-EcoRI and BEL-7402-hTR- BamH I cells), blank control group (BEL-7402 cells). Adhesion and invasion assays were used to detect the adhesion and invasion ability of BEL-7402 cells after antisense hTR transfection. The expression of integrinβ1, INTβ1 and E-cad was detected by immunocytochemistry Gelatin zymography was used to detect the changes of matrix metalloproteinase (MMP) -2 and -9 activity. Results: The adhesion and invasion ability of BEL-7402-hTR-BamHI cells in the antisense hTR transfected group was significantly lower than that in the blank control group and the positive hTR transfection group (0.204 ± 0.029 vs 0.505 ± 0.037 and 0.465 ± 0.029, (P <0.05). The expression of E-cad was significantly higher in the control group (34.80 ± 3.19vs71.47 ± 5.15,69.87 ± 3.11, P <0.05) vs0.124 ± 0.018,0.134 ± 0.016, P <0.05), the activity of MMP-2 and MMP-9 were significantly inhibited (2 054.22 ± 138.52 vs3 105.56 ± 329.60, 2 923.22 ± 269.08, 846.33 ± 104.66 vs 538.89 ± 122.85, 1 453.33 ± 126.35, all P <0.05). CONCLUSION: Human antisense telomerase RNA can significantly reduce the adhesion and invasion ability of BEL-7402 hepatocellular carcinoma cells. The mechanism may be related to up-regulating the expression of E-cad, down-regulating the expression of INTβ1 and inhibiting the activity of MMP-2 and MMP-9 related.