磷脂酶D2促进结直肠癌细胞生长的体内实验研究

Experimental study on the effect of phospholipase D2 on the growth of colorectal cancer cells in vivo

目的利用体内实验探讨磷脂酶D2(PLD2)基因表达对结直肠癌细胞生长的影响及可能的机制。方法建立裸鼠结直肠癌(SW480和SW620)移植瘤模型,待肿瘤大小达50~70 mm^(3)时,将携带PLD2-siRNA的腺病毒、空腺病毒载体或生理盐水向瘤体内多方向注射,并依此将2种细胞株均分为PLD2干扰组、空载组及正常组;实时荧光定量PCR和蛋白质印迹法检测瘤体内PLD2的mRNA和蛋白表达水平;根据实验期间肿瘤的生长情况,计算肿瘤近似体积,绘制瘤体生长曲线和比较2种细胞株各组的瘤体质量;蛋白质印迹法检测瘤体内c-Myc和Bcl-2的蛋白表达水平。结果携带PLD2-siRNA腺病毒载体使瘤体内PLD2 mRNA和蛋白表达水平降低(P<0.001),干扰降低PLD2基因表达导致结直肠癌细胞的生长受抑制(P<0.001),同时c-Myc蛋白的表达降低(P<0.001),而Bcl-2的表达升高(P<0.001)。结论干扰降低PLD2基因表达导致结直肠癌细胞生长受抑制,其可能与c-Myc表达降低和Bcl-2表达升高相关,提示PLD2可能成为结直肠癌治疗的有效靶点。

Objective To explore the effect and possible mechanism of phospholipase D2(PLD2)gene expression on the growth of colorectal cancer cells from in vivo experiments.Methods The nude mice model of colorectal cancer(SW480 and SW620)was established.When tumor size reached 50-70 mm^(3),adenovirus carrying human PLD2-siRNA,or no virus was injected into the tumor in multiple directions.The cell lines were divided into the PLD2 interference group,mock-vehicle group and normal group.The real-time quantitative PCR and western blotting were used to detect the mRNA and protein expression level of the PLD2 in tumors.According to the maximum transverse diameter(L)and maximum longitudinal diameter(S)of the tumor were measured during the experiment,and the tumor growth curve was calculated after calculating the tumor approximate volume.The tumor mass of each cell group was compared.Western blotting was used to detect the expression levels of the c-Myc and Bcl-2 proteins in tumor.Results The PLD2-siRNA led to the decrease in mRNA and protein expression of the PLD2 in tumors(P<0.001).And the interference with reduced PLD2 gene expression led to the growth of colorectal cancer cells inhibit(P<0.001).At the same time,the expression of c-Myc protein was decreased(P<0.001),while the expression of Bcl-2 was increased(P<0.001).Conclusion The interfering PLD2 gene expression may lead to the inhibition of the growth of colorectal cancer cells,which may be closely related to the decrease of the c-Myc expression and increase of Bcl-2 expression.It′s suggested that the PLD2 may be an effective target for the colorectal cancer treatment.