过表达HepaCAM联合谷氨酰胺剥夺抑制前列腺癌细胞的增殖

Overexpression of HepaCAM Combined with Glutamine Deprivation Inhibits the Proliferation of Prostate Cancer Cells

该研究旨在探讨过表达肝细胞黏附分子(hepatocyte cell adhesion molecule,HepaCAM)联合谷氨酰胺(glutamine,Gln)剥夺对前列腺癌(prostae cancer,PCa)细胞谷氨酰胺代谢及增殖的影响。采用Ad-HepaCAM腺病毒感染前列腺癌细胞株PC3、LNCaP。克隆形成实验及MTT实验检测细胞的克隆形成率及增殖活性。qRT-PCR、Western blot分别检测PC3、LNCaP细胞中谷氨酰胺酶(glutaminase,GLS)、溶质载体家族I成员V(solute carrier family 1 member 5,SLC1A5)、MYC癌基因家族(可编码c-MYC蛋白分子)以及细胞周期蛋白D1(cyclin D1)、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的表达情况。结果显示,单独过表达HepaCAM与剥夺Gln均可抑制PC3、LNCaP细胞的增殖能力,两者联用效果更佳。qRT-PCR和Western blot显示,与对照组(+Gln组)相比,剥夺谷氨酰胺组(-Gln组)细胞中的GLS、SLC1A5、MYC表达呈应激抵抗式上调,过表达HepaCAM后上述基因表达重新受到抑制。该实验证明,过表达HepaCAM与Gln剥夺联用可更显著地抑制前列腺癌的增殖能力,同时,过表达HepaCAM可在一定程度上抑制前列腺癌细胞的谷氨酰胺代谢重编程。

The purpose of this study was to investigate the effect of HepaCAM(hepatecyte cell adhesion molecule)combined with Gln(glutamine)deprivation on glutamine metabolism and proliferation of PCa(prostate cancer)cells.Prostate cancer cell lines PC3 and LNCaP were infected with Ad-HepaCAM adenovirus.Clone formation experiment and MTT assay were used to detect the clonogenesis rate and proliferation activity.The expression of GLS(glutaminase),SLC1A5(solute carrier family I member V),MYC oncogene family,cyclin D1 and PCNA(proliferating cell nuclear antigen)were detected by qRT-PCR and Western blot.The results showed that overexpression of HepaCAM and deprivation of Gln could inhibit the proliferation of PC3 and LNCaP cells,and the combination of them worked better.qRT-PCR and Western blot showed that compared with the+Gln group(control group),the expression of GLS,SLC1A5 and MYC in the-Gln group(glutamine deprivation group)was up-regulated in a stress-resistant manner,and above the genes were inhibited again after overexpression of HepaCAM.This experiment proves that the combination of overexpression of HepaCAM and Gln deprivation can significantly inhibit the proliferation of prostate cancer,and overexpression of HepaCAM can inhibit the reprogramming of glutamine metabolism in prostate cancer cells to some extent.