| 雷志杰,张 莹,田苗苗,岳俊涛,夏云龙,胡思隽,吴开春.TGM2对胃癌细胞BGC-823增殖、迁移和侵袭能力的影响[J].,2018,(10):1816-1822 |
| TGM2对胃癌细胞BGC-823增殖、迁移和侵袭能力的影响 |
| The Role of TGM2 on Cell Proliferation, Migration and Invasion of BGC-823 Cell in Gastric Cancer |
| 投稿时间：2018-02-05 修订日期：2018-03-02 |
| DOI：10.13241/j.cnki.pmb.2018.10.004 |
| 中文关键词: 胃癌 TGM2 细胞增殖 肿瘤转移 |
| 英文关键词: Gastric cancer TGM2 Cell proliferation Tumor metastasis |
| 基金项目:国家自然科学基金项目（81421003，81627807，91542000，81672702，81701750） |
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| 中文摘要: |
| 摘要 目的：转谷氨酰胺酶-2（transglutaminase-2, TGM2）是一种多功能的蛋白，在乳腺癌，胰腺癌，黑色素瘤和前列腺癌等肿瘤中高表达，而且和肿瘤的增殖和转移密切相关。本研究旨在探索TGM2对胃癌细胞BGC-823的增殖、迁移和侵袭能力的影响。方法：qRT-PCR、蛋白质印迹及免疫组化检测TGM2在胃癌细胞及组织中的表达情况；慢病毒转染胃癌细胞BGC-823以干扰TGM2的表达，荧光显微镜下观察转染效率，蛋白质印迹检测TGM2表达干扰效果；CCK-8法检测TGM2下调对BGC-823增殖能力的影响；Transwell法检测TGM2下调对BGC-823的迁移和侵袭能力影响；蛋白质印迹检测TGM2下调对转移和凋亡相关蛋白表达的影响。结果：TGM2在胃癌细胞和组织中均高表达，P<0.01（qRT-PCR），P<0.001（免疫组化）；转染慢病毒后，荧光显微镜下观察结果显示转染效率超过90 %，蛋白质印迹结果显示实验组shTGM2-1的干扰效果最好，TGM2明显下调，差异具有统计学意义，P<0.0001；CCK-8结果表明TGM2下调后，从细胞铺板第2d开始，实验组（shTGM2-1）的细胞增殖倍数明显下降，P<0.05(2d)，P<0.01(3d)，P<0.001(4d)，P<0.01(5d)；Transwell结果显示，TGM2下调后，BGC-823的迁移和侵袭能力明显减弱，P<0.001(迁移)，P<0.01(侵袭)；蛋白质印迹结果显示，TGM2下调后，NF-κB和Bcl-2表达下调，而Bax表达上调，P<0.05(NF-κB)、P<0.001 (Bcl-2)和P<0.0001(Bax)。结论：TGM2下调能抑制胃癌细胞BGC-823的增殖、迁移和侵袭能力，并和NF-κB、Bcl-2的下调以及和Bax的表达水平上调有关，为胃癌的临床治疗提供了新靶点。 |
| 英文摘要: |
| ABSTRACT Objective: Transglutaminase-2 (TGM2) is a multifunctional protein that is overexpressed in tumors such as breast, pancreatic, melanoma and prostate cancer, and is closely related to tumor proliferation and metastasis. The purpose of this study was to explore the effect of TGM2 on the proliferation, migration and invasion of gastric cancer cell line BGC-823. Methods: The expression of TGM2 in gastric cancer cells and tissues was detected by qRT-PCR, Western blotting and immunohistochemistry. The lentivirus was used to transfect BGC-823 to down-regulate the expression of TGM2 and the transfection efficiency was observed by fluorescence mi- croscopy and Western blotting. The effect of TGM2 down-regulation on the proliferation of BGC-823 cells was detected by CCK-8 as- say. The effect of down-regulation of TGM2 on the migration and invasion ability of BGC-823 was detected by Transwell assay. The down-regulation of TGM2 on the metastasis and apoptosis-related protein expression was detected by Western blotting. Results: TGM2 was highly expressed in gastric cancer cells (P<0.01) and tissues (P<0.001). The results of fluorescence mi- croscopy showed that trans- fection efficiency was more than 90 %. The results of Western blotting indicated that the interference effect of shTGM2-1group was best obvious(P<0.0001). The results of CCK-8 showed that after TGM2 down-regula- tion, from the first day, cell proliferation in the experi- mental group (shTGM2-1) was significantly decreased. P<0.05(2d), P<0.01(3d)，P<0.001(4d) and P<0.01(5d). Transwell assays showed that the migration (P<0.001) and invasion (P<0.01) ability of BGC-823 were sup- pressed after TGM2 down-regulation. Western blotting showed that, expression of NF-κB (P<0.05) and Bcl-2 (P<0.001) were down-regulated while Bax (P<0.0001) was up-regulated in BGC-823 when TGM2 was down- regulated. Conclusion: The proliferation, migration and invasion abilities of gastric cancer cell BGC-823 are inhibited significantly after downregulation of TGM2, which is related to the down-regulation expression of NF-κB, Bcl-2 and the up-regulation expression of Bax. These results provide a new target for the clinical treatment of gastric cancer. |
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