HMGI-C suppressing induces P53/caspase9 axis to regulate apoptosis in breast adenocarcinoma cells
HMGI-C suppressing induces P53/caspase9 axis to regulate apoptosis in breast adenocarcinoma cells
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DOI：
10.1080/15384101.2016.1190892
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发表时间：
2016-01-01
期刊：
影响因子：
4.3
通讯作者：
Baradaran, Behzad
中科院分区：
文献类型：
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作者：
Mansoori, Behzad;Mohammadi, Ali;Baradaran, Behzad
目的：高迁移率族蛋白异构体(HMGI-C)是高迁移率族AT-钩蛋白(HMGA)家族中的一员，它是一类小的非组蛋白染色体蛋白，可以调节大量基因的转录。全基因组研究显示，HMGI-C基因在许多人类癌症中上调，这表明HMGI-C可能在各种肿瘤的发展中发挥关键作用。然而，HMGI-C在乳腺癌中的确切作用尚不清楚。方法：采用定量逆转录聚合酶链式反应(QRT-PCR)方法检测HMGI-C在乳腺癌组织和癌旁正常组织中的表达。用四甲基偶氮唑盐比色法检测HMGI-C siRNA对乳腺癌细胞的杀伤作用。用实时定量聚合酶链式反应和Western blotting分别检测HMGI-C的相对mRNA和蛋白水平。采用原位末端标记法和Annexin-V/PI法检测细胞凋亡，蛋白凝胶印迹法检测p53、caspase3、9、8和bcl2蛋白表达，QRT-PCR法检测miR34a、let-7a基因表达。用碘化丙啶DNA染色流式细胞仪分析细胞周期。结果：HMGA2在乳腺癌组织和边缘正常组织中的过度表达差异有高度统计学意义(P<；0.0001)。HMGI-C siRNA在转染后48小时内显著降低mRNA和蛋白质表达水平，且呈剂量依赖关系。我们观察到HMGI-C在体外通过诱导miR34a和细胞周期停滞而通过线粒体途径显著诱导MDA-MB-468细胞的凋亡。结论：HMGI-C可能在乳腺癌的发生发展中起重要作用。在这里，我们介绍了HMGI-C作为一种潜在的治疗靶点，以触发人乳腺腺癌的细胞凋亡和细胞周期停滞。因此，HMGI-C siRNA可能是治疗乳腺癌的有效佐剂。
Purpose: The HMGI-C (high mobility group protein isoform I-C) protein is a member of the high-mobility group AT-hook (HMGA) family of small non-histone chromosomal proteins that can modulate transcription of an ample number of genes. Genome-wide studies reveal upregulation of the HMGI-C gene in many human cancers, which suggests that HMGI-C might play a critical role in the progression of various tumors. However, the exact role of HMGI-C in breast adenocarcinoma has not been made clear. Methods: HMGI-C mRNA expression in breast cancer samples and marginal normal tissues was characterized using qRT-PCR. The cytotoxic effects of HMGI-C siRNA on breast adenocarcinoma cells were determined using MTT assay. Relative HMGI-C mRNA and protein levels were measured by quantitative real-time PCR and western blotting, respectively. Apoptosis detection was done using TUNEL and Annexin-V/PI assays, P53, caspase 3, 9, 8 and Bcl2 proteins evaluated by protein gel blot and miR34a, Let-7a genes investigates by QRT-PCR assay. Cell cycle was analyzed by flow cytometry assay using propidium iodide DNA staining. Results: An overexpression of HMGA2 was revealed with highly statistically significant differences between breast cancer samples and marginal normal tissues (P < 0.0001). HMGI-C siRNA significantly reduced both mRNA and protein expression levels in a 48-hour period after transfection and in a dose-dependent manner. We observed that the knockdown of HMGI-C led to the significant induction of apoptosis via mitochondrial pathway by inducing miR34a and cell cycle arrest in MDA-MB-468 cells in vitro. Conclusions: These results propose that HMGI-C might play a critical role in the progression of breast adenocarcinoma. Here we introduced HMGI-C as a potential therapeutic target for trigger apoptosis and cell cycle arrest in human breast adenocarcinoma. Therefore HMGI-C siRNA may be an effective adjuvant in human breast adenocarcinoma.