HDGF的PWWP结构域改变对肿瘤细胞体外及体内增殖的影响

Effects of Mutant PWWP Domain of HDGF on the Proliferation of Tumor Cells in Vitro and in Vivo

目的:研究肝癌衍生生长因子(Hepatoma-derived growth factor,HDGF)PWWP结构域(PWWP domain)改变对肿瘤细胞体外及体内增殖的影响。方法:构建HDGF的PWWP结构域突变体P24A,利用慢病毒感染细胞筛选稳定细胞系。采用CCK-8法和软琼脂克隆形成实验检测细胞的增殖情况。通过裸鼠皮下成瘤实验检测移植瘤的形成情况。结果:在肝癌Hep G2和结直肠癌DLD1稳定细胞株中,CCK-8法检测结果显示突变体P24A对细胞生长的抑制作用呈时间依赖性,其OD值在24、48、72和96 h均明显低于HDGF稳定细胞株(均P<0.001)。克隆形成实验结果显示P24A组克隆数目明显小于HDGF组(P<0.01)。异种移植瘤动物模型则证明P24A细胞株的瘤块生长速度(P<0.001,P<0.01,P<0.01),瘤块大小及体积(P<0.01)均明显低于HDGF细胞株。结论:PWWP结构域改变可能抑制HDGF发挥促进细胞增殖的作用。

Objective： To identify the effect of mutant PWWP domain of Hepatoma-derived growth factor（HDGF） on the proliferation of tumor cells in vitro and in vivo. Methods： The mutated PWWP domain of HDGF was constructed, named P24 A. Cells were infected with lentivirus supernatants to screen stable cell lines. CCK-8 assay and soft agar colony formation assay were performed to detect the proliferation in vitro. Cells stably expressing HDGF and P24 A were injected subcutaneously into the BALB/c nude mice to verify the effect on the proliferation in vivo. Results： The CCK-8 assay showed that the inhibitory effect of mutant P24 A on cell growth was time-dependent（24 h, P〈0.001; 48 h, P〈0.001; 72 h, P〈0.001; 96 h, P〈0.001） compared with that of HDGF in liver cancer stable cell lines Hep G2 and colorectal cancer stable cell lines DLD1. And so was the colony formation ability（P〈0.01）. Animal models of subcutaneous xenograft in BALB/c nude mice demonstrated that the growth rate（P〈0.001, P〈0.01, P〈0.01, respectively）, tumor size and volume（P〈0.01） of P24 A cell line were significantly lower than that of HDGF. Conclusions： The mutant PWWP domain may inhibit the role of HDGF in promoting cell proliferation.