miR-506-3p通过上调SDAD1促进结直肠癌细胞HT-29的凋亡

miR-506-3p induces apoptosis of colon cancer cell HT-29 by upregulating SDAD1

[目的]探讨miR-506-3p是否靶向SDAD1并调控结直肠癌细胞HT-29凋亡的发生。[方法]通过TargetScan在线分析miR-506-3p与SDAD1的匹配度,利用双荧光素酶报告系统检测miR-506-3p与SDAD1的结合。进一步过表达与干扰miR-506-3p,通过Western Blot法检测SDAD1及凋亡标志蛋白的表达情况,通过Annexin-V染色检测结直肠癌细胞HT-29的凋亡水平。[结果]miR-506-3p可以与SDAD1的3′端非编码689-692区域靶向结合。过表达miR-506-3p后,SDAD1与凋亡标志蛋白Cleaved-caspase 3和Cleaved-caspase 9的表达量上升,结直肠癌细胞HT-29的凋亡水平显著上升。干扰miR-506-3p后,SDAD1、凋亡标志蛋白Cleaved-caspase 3和Cleaved-caspase 9的表达量下降,结直肠癌细胞HT-29的凋亡水平显著下降。[结论]miR-506-3p可以靶向结合SDAD1从而上调其表达,而SDAD1是结直肠增殖、凋亡相关基因,miR-506-3p通过上调SDAD1促进结直肠癌细胞HT-29的凋亡。提示miR-506-3p的表达可能作为SDAD1靶向RNA反映结直肠癌的增殖,与结直肠癌的发生、进展相关,进一步探究miR-506-3p抑制剂在结直肠癌患者治疗方面的应用具有良好的临床应用前景。

[Objective]To investigate whether miR-506-3 p could regulate the apoptosis of colon cancer cell HT-29 by targeting SDAD1.[Method] TargetScan on-line analyzed whether miR-506-3 p matched SDAD1.Secondly,the dual luciferase reporter system was used to detect whether miR-506-3 p targeted SDAD1.In the case of overexpression or knockdown of miR-506-3 p,the expression of SDAD1 and apoptosis marker proteins was detected by Western Blot.Annexin-V staining was used to detect the apoptosis level of colorectal cancer HT-29.[Result]The 3 non-coding 689-692 region of SDAD1 could be targeted and bound by miR-506-3 p.After overexpression of miR-506-3 p,the expression of SDAD1,apoptosis marker proteins Cleaved-caspase3 and Cleaved-caspase9 increased,and the apoptosis level of colorectal cancer cell HT-29 increased significantly.After knocking down miR-506-3 p,it was found that the expression of SDAD1,the apoptosis marker proteins Cleaved-caspase3 and Cleaved-caspase9 decreased,and the apoptosis level of colorectal cancer cell HT-29 decreased significantly.[Conclusion]miR-506-3 p could bind SDAD1 to up-regulate its expression.SDAD1 is a gene related to colorectal proliferation and apoptosis.Thus,miR-506-3 p may promote the apoptosis of HT-29 in colorectal cancer cells by up-regulating SDAD1.It is suggested that the expression of miR-506-3 p may reflect the proliferation of colorectal cancer as SDAD1 targeting RNA,which indicated the occurrence and progression of colorectal cancer.Further exploration of the application of miR-506-3 p inhibitor in the treatment of colorectal cancer patients has a good clinical application prospect.