细胞自噬与结肠癌Lovo细胞迁移和侵袭的相关性研究

The relationship of cell autophagy and Lovo cell migration and invasion

目的探讨不同自噬状态对结直肠癌Lovo细胞迁移和侵袭能力的影响。方法将Lovo细胞分为自噬增强组、正常细胞组和3-甲基嘌呤自噬抑制组。用激光共聚焦显微镜观察绿色荧光颗粒,用Q-PCR检测Beclin1 mRNA表达水平,透射电镜观察自噬溶酶体,Transwell实验评价Lovo细胞迁移与侵袭能力。结果绿色荧光颗粒数以自噬增强组最多,其次为正常细胞组,而自噬抑制组最少。Beclin1 mRNA表达量自噬增强组为(1.23±0.12)个,正常细胞组为(1±0.13)个,自噬抑制组为(0.98±0.1)个。自噬增强组可见大量的自噬溶酶体,明显多于正常细胞组和自噬抑制组。迁移实验细胞计数:自噬增强组高于正常细胞组(138.0±16.7与90.7±12.9,P=0.026)和自噬抑制组(138.0±16.7与92.7±26.7,P=0.030)。侵袭实验细胞计数:自噬增强组高于正常细胞组(147.0±13.0与99.0±20.5,P=0.028)和自噬抑制组(147.0±13.0与95.7±25.6,P=0.021)。结论结肠癌Lovo细胞自噬增强促进肿瘤细胞迁移和浸润,可能是局部浸润和远处转移的机制之一。

Objective To investigate the influence of autophagy on migration and invasion in colon cancer cells. Methods The Lovo cells of colon cancer were classified as autophagy-enhanced group, normal cell group, and 3-Methyladenine autophagy-inhibited group. Laser canning eonfocal microscope was used to identify the green fluorescence particles. Beelinl mRNA expressive level was measured with Q-PCR. Transmission electron microscope was employed to observe the autolysosome. Transwell chamber was applied to evaluate the migration and invasion of Lovo cell. Results There were more green fluorescence particles in the autophagy-enhanced group than the normal cell group and the autophagy-inhibited group. The Beclin 1 mRNA expressive level was 1.23 ±0.12 in the autophagy-enhaneed group,while 1 ±0.13 in the normal cell group and 0.98 ± 0. 1 in the autophagy-inhibited group. There were the most autolysosomes in autophagyenhanced group of all groups. The migration experiment showed the mean number of Lovo cell in the autophagy-enhanced group（ 138.0± 16.7 ）were more than the normal Lovo cell group （90.7 ± 12.9, P = 0.026 ） and the autophagy-inhibited group （ 92.7 ± 26.7, P = 0. 030 ）. The invasion experiment found the mean number of Lovo cell in autophagy-enhanced group （ 147.0 ± 13.0 ）were also more than the normal Lovo cell group （ 99. 0 ±20. 5, P = 0. 028 ） and the autophagy-inhibited group （ 95.7 ± 25.6, P = 0. 021 ）. Conclusions Autophagy enhancement could promote the migrating and invasive ability of Lovo cell. And that might be an important mechanism of invasion and metastasis in malignant tumors.