抑癌基因PTEN的表达对大肠癌细胞转移侵袭能力的影响

The influence of PTEN protein expression on the invasive and sdhesive ability of colorectal cancer cell

目的 探讨抑癌基因PTEN的表达大肠癌细胞转移侵袭能力的影响。方法 1.利用westernblot法检测不同转移潜能的大肠癌细胞系内PTEN蛋白的表达水平 ,说明PTEN蛋白的表达对大肠癌细胞转移潜能的影响 ,2 .用脂质体作载体 ,将PTEN基因转染大肠癌细胞株LOVO后 ,采用计数细胞悬液加到粘附底物后 2 0min和 12 0min的细胞贴壁数用以测定细胞粘附能力 ,采用Costar的浸润小室检测PTEN基因转染前后细胞的浸润能力。结果 1.转移潜能高的LOVO细胞PTEN的表达量显著低于转移潜能较低的HT 2 9,LS 174T ,2 .未转染细胞 (LOVO)、转染 pcDNA3.0 PTEN的细胞 (LOVO/pcDNA3.0 PTEN)在特异性粘附底物 (Laminin)上 2 0min时贴壁率分别为 18.6 %± 1.4 %和 13.9%± 0 .4 8% (P <0 .0 5 ) ,12 0min时贴壁率分别为 71.2 %± 2 .5 %和 5 6 .0 %± 1.6 % (P <0 .0 5 ) ,3.采用Costar的浸润小室对LOVO、LOVO/pcDNA3.0 PTEN细胞的浸润能力分析结果显示 :细胞悬液静置培养 6h后 ,对照细胞LOVO浸润穿透多聚碳膜的细胞数为 11.7± 1.74个 ,LO VO/pcDNA3.0 PTEN细胞穿透多聚碳膜的细胞数为 7.5± 1.5 8个 (P <0 .0 5 )。 结论 在肿瘤细胞内抑癌基因PTEN的表达与大肠癌的转移侵袭行为密切相关。

Objective To investigate the influence of PTEN protein expression on the invasive and adhesive ability of colorectal cancer cell. Methods 1. Protein expression of PTEN were tested in three colorectal cancer cell line with different metastasic ability by western-blot.2.The human PTEN cDNA containing eukaryotic expression vectors pcDNA3.0 was transferred into human colorectal cancer cell line LOVO, to investigate the changes of invasive and adhesive ability of LOVO by Boyden chamber and a microscope slide combined with Laminin. Results 1. The content of PTEN protein in LOVO, which had a higher ability of mestastase lower than that in HT-29,LS-174T. 2.The adhesive rate of LOVO and LOVO/pcDNA3.0-PTEN to laminin at 20mins is 18.6% ±1.4% and 13.9% ±0.48%(P<0.05),at 120mins is 71.2% ±2.5% and 56.0% ±1.6% (P<0.05)respectively,3. The average number of LOVO and LOVO/pcDNA3.0-PTEN,whitch could transit through Boyden chamber was 11.7±1.74 and 7.5 ±1.58个(P<0.05), Conclusion The expression of PTEN may play an important role in the metastase process of colorectal cancer.