结肠癌中HRH3表达对细胞增殖的影响

Effects of the expression of HRH3 on cell growth of colon cancer cells

目的探讨组胺受体3(histamine receptor 3,HRH3)对结肠癌细胞恶性增殖的影响及其相关信号通路。方法以结肠癌Ls174T细胞和SW480为研究对象,研究HRH3对其生长增殖的影响。通过转染siRNA片段沉默HRH3基因表达。CCK-8检测细胞生长,EdU检测细胞增殖活性,实时荧光定量PCR和Western blot检测相关信号通路分子改变。结果转染siRNA片段可有效沉默HRH3表达。实时荧光定量PCR和Western blot结果显示,转染si-HRH3后,HRH3 mRNA和蛋白表达均显著降低(P均<0.05)。CCK-8实验结果显示,沉默HRH3后,细胞生长明显减慢(P均<0.05)。EdU实验结果显示,沉默HRH3后,细胞增殖活性显著降低(P均<0.05)。机制研究表明,沉默HRH3后,c-Myc表达显著降低、CDKN1A表达显著升高(P均<0.05)。结论HRH3可能通过调控c-Myc和CDKN1A转录表达促进结肠癌细胞的恶性增殖。

Purpose To explore the effect of histamine receptor 3(HRH3)on the malignant proliferation of colon cancer cells and related signal pathways.Methods siRNA specially targeted to HRH3 was applied to silence the expression of HRH3 in colon cancer cells.Cell growth and cell proliferation were detected by CCK-8 and EdU assay.The mRNA and protein expression of the related signal pathways molecules were detected by real-time PCR and Western blot.Results The mRNA and protein level of HRH3 was significantly decreased after transfection of si-HRH3 by real-time PCR and Western blot,respectively(all P<0.05).Moreover,our data showed that HRH3 silencing significantly attenuated the cell growth of colon cancer cells via CCK-8 assay(all P<0.05).Furthermore,EdU assay showed that HRH3 silencing remarkably reduced the cell proliferation activity of colon cancer cells(all P<0.05).For the mechanism,their results indicated that the expression of c-Myc was significantly decreased,and the expression of CDKN1A was significantly increased after silencing HRH3 in colon cancer cells(all P<0.05).Conclusion HRH3 promotes the malignancy growth via regulating the expression of c-Myc and CDKN1A in colon cancer.