BACH1 silencing by siRNA inhibits migration of HT-29 colon cancer cells through reduction of metastasis-related genes.
BACH1 silencing by siRNA inhibits migration of HT-29 colon cancer cells through reduction of metastasis-related genes.
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DOI：
10.1016/j.biopha.2016.09.021
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发表时间：
2016-12-01
期刊：
影响因子：
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通讯作者：
Baradaran, Behzad
中科院分区：
文献类型：
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作者：
Davudian, Sadaf;Shajari, Neda;Baradaran, Behzad
背景：转移到远处器官是许多肿瘤细胞的标志。BACH 1（BTB and CNC homology 1）是一种促进乳腺癌细胞迁移和侵袭的转录因子。BACH 1表达及其靶基因与临床样本的转移可能性密切相关，BACH 1减少导致转移中有意义的耗竭。BACH 1在结肠癌中的作用的评估仍然是难以捉摸的。方法：采用定量RT-PCR（qRT-PCR）方法检测siRNA敲除结肠癌HT-29细胞后BACH 1及相关转移基因的表达。Western blot检测蛋白水平。MTT法检测转染BACH 1 siRNA后细胞活力的变化。Sjuch-wound运动实验检测了BACH 1沉默后HT-29细胞的迁移能力。结果：利用高转移性HT-29结肠癌细胞系，通过siRNA敲低BACH 1的方法检测了BACH 1的抑制作用。定量RT-PCR和Western blot分析显示，转染后HT 29细胞BACH 1 mRNA和蛋白的表达水平均受到明显抑制。相反，BACH 1表达增加了迁移。此外，CXCR 4和MMP 1的表达水平下降后，BACH 1敲低在HT-29 cells.CONCLUSION：我们的结果表明，BACH 1下调在HT-29 CRC细胞中没有影响细胞生长，但通过降低转移相关基因的表达抑制细胞迁移。总的来说，这些结果表明BACH 1可能在结肠癌中起致癌驱动作用，并可能代表CRC治疗的基因治疗的潜在靶点。
BACKGROUND: Metastasis to distant organs is a hallmark of many tumor cells. BACH1 (BTB and CNC homology 1) is a transcriptional factor which promotes the migration and invasion of breast cancer cells. BACH1 expression and its target genes are intimately associated with the metastasis possibility of clinical samples, and BACH1 reduction leads to meaningful depletion in metastasis. The evaluation of BACH1 role in colon cancer remains elusive. This study seeks to further investigate the role of BACH1 in colon cancer cells.METHODS: Quantitative RT-PCR (qRT-PCR) was used to detect BACH1 expression and other related metastatic genes following siRNA knockdown in colon cancer HT-29 cells. And the protein level assessed by Western blot. MTT assay was to measure the changed cell viability after BACH1 siRNA transfection. Scratch-wound motility assays measured capacity of tumor cell migration of HT-29 cells after BACH1 silencing.RESULTS: The inhibitory effect of BACH1 was performed by siRNA knockdown using highly metastatic HT-29 colon cell lines. Quantitative RT-PCR and Western blot analysis revealed that the expression levels of BACH1 mRNA and protein in HT29 cells were significantly suppressed after transfection. Conversely, the BACH1 expression increased migration. Also the CXCR4 and MMP1 expression levels decreased following BACH1 knockdown in HT-29 cells.CONCLUSION: Our results indicated that BACH1 down-regulation in HT29 CRC cells had no effect on cell growth but did inhibit cell migration by decreasing metastasis-related genes expression. Collectively, these results suggest that BACH1 may function as an oncogenic driver in colon cancer and may represent as a potential target of gene therapy for CRC treatment.