反义端粒酶RNA对肝癌细胞黏附和侵袭的影响

Effect of antisense telomerase RNA on adhesion and invasion of hepatocellular carcinoma cells

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 study the effect of antisense human telomerase RNA（hTR） on adhesion and invasion of hepatocellular carcinoma（HCC） cell line BEL-7402 and its mechanism.Methods： Our previous research had successfully transfected sense or antisense hTR gene into BEL-7402 cells.This study they were divided into 3 groups： the sense hTR transfected group（BEL-7402-hTR-EcoRI）,the antisense hTR transfected group（BEL-7402-hTR-BamHI） and the blank control group（BEL-7402）.The adhesion and invasion of BEL-7402 cells after antisense hTR transfection were observed with adhesion and invasive assay,respectively.The expressions of integrin β1（INTβ1） and epithelial cadherin（E-cad） were detected by immunocytochemistry,and galatin zymography was used to measure the activities of matrix metalloproteinase-2（MMP-2） and MMP-9.Results： The adhesion and invasion abilities of antisense hTR transfected BEL-7402 cells were significantly weaker than those of the blank control and sense hTR transfected groups（0.204±0.029 vs 0.505±0.037,0.465±0.029;34.80±3.19 vs 71.47±5.15,69.87±3.11;all P〈0.05）.Meanwhile,the expression of INTβ1 was also significantly decreased（0.153 3±0.015 6 vs 0.385±0.008,0.375±0.014 4,P〈0.05）,and E-cad was significantly increased（0.209±0.020 vs 0.124±0.018,0.134±0.016;P〈0.05） in the antisense hTR transfected group.The activities of MMP-2 and MMP-9 were significantly inhibited in the antisense hTR transfected group（2 054.22±138.52 vs 3 105.56±329.60,2 923.22±269.08;846.33±104.66 vs 1 538.89±122.85,1 453.33±126.35;all P〈0.05）.Conclusion： Antisense hTR can inhibit the adhesion and invasion of hepatocellular carcinoma BEL-7402 cells,which may be related to the increase of E-cad expression,decrease of INTβ1 expression,and decrease of MMP-2 and MMP-9 activities.