Davudian Sadaf, Shajari Neda, Kazemi Tohid, Mansoori Behzad, Salehi Shima, Mohammadi Ali, Shanehbandi Dariush, Shahgoli Vahid Khaze, Asadi Milad, Baradaran Behzad
Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.
Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Biomed Pharmacother. 2016 Dec;84:191-198. doi: 10.1016/j.biopha.2016.09.021. Epub 2016 Sep 19.
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.
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.
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.
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.
远处器官转移是许多肿瘤细胞的一个标志。BACH1(BTB和CNC同源物1)是一种转录因子,可促进乳腺癌细胞的迁移和侵袭。BACH1的表达及其靶基因与临床样本的转移可能性密切相关,BACH1的减少会导致转移的显著减少。BACH1在结肠癌中的作用评估仍不明确。本研究旨在进一步探讨BACH1在结肠癌细胞中的作用。
使用定量RT-PCR(qRT-PCR)检测结肠癌HT-29细胞中siRNA敲低后BACH1的表达及其他相关转移基因。通过蛋白质印迹法评估蛋白质水平。MTT法用于测量BACH1 siRNA转染后细胞活力的变化。划痕伤口运动试验测量BACH1沉默后HT-29细胞的肿瘤细胞迁移能力。
使用高转移性HT-29结肠癌细胞系通过siRNA敲低来实现对BACH1的抑制作用。定量RT-PCR和蛋白质印迹分析显示,转染后HT29细胞中BACH1 mRNA和蛋白质的表达水平显著受到抑制。相反,BACH1的表达增加了迁移。在HT-29细胞中BACH1敲低后,CXCR4和MMP1的表达水平也降低。
我们的结果表明,HT29结直肠癌细胞中BACH1的下调对细胞生长没有影响,但通过降低转移相关基因的表达确实抑制了细胞迁移。总体而言,这些结果表明BACH1可能在结肠癌中作为致癌驱动因子发挥作用,并且可能代表结直肠癌治疗的基因治疗潜在靶点。