Zhou Xiao-Na, Li Guang-Ming, Xu Ying-Chen, Zhao Tuan-Jie, Wu Ji-Xiang
Department of General Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China.
Department of General Surgery, Beijing Er Long Lu Hospital, Beijing 100032, China.
Chin Med J (Engl). 2016 Nov 5;129(21):2623-2629. doi: 10.4103/0366-6999.192775.
Decoy receptor 3 (DcR3) binds to Fas ligand (FasL) and inhibits FasL-induced apoptosis. The receptor is overexpressed in hepatocellular carcinoma (HCC), and it is associated with the growth and metastatic spread of tumors. DcR3 holds promises as a new target for the treatment of HCC, but little is known regarding the molecular mechanisms underlying the oncogenic properties of DcR3. The present work, therefore, examined the role of DcR3 in regulating the growth and invasive property of liver cancer cell HepG2.
HepG2 cells were stably transfected with lentivirus-based short hairpin RNA vector targeting DcR3. After the knockdown of DcR3 was confirmed, cell proliferation, clone formation, ability of migrating across transwell membrane, and wound healing were assessed in vitro. Matrix metalloproteinase-9 (MMP 9) and vascular epithelial growth factor (VEGF)-C and D expressions of the DcR3 knockdown were also studied. Comparisons between multiple groups were done using one-way analysis of variance (ANOVA), while pairwise comparisons were performed using Student's t test. P< 0.05 was regarded statistically significant.
DcR3 was overexpressed in HepG2 compared to other HCC cell lines and normal hepatocyte Lo-2. Stable knockdown of DcR3 slowed down the growth of HepG2 (P < 0.05) and reduced the number of clones formed by 50% compared to those without DcR3 knockdown (P < 0.05). The knockdown also reduced the migration of HepG2 across transwell matrix membrane by five folds compared to the control (P < 0.05) and suppressed the closure of scratch wound (P < 0.05). In addition, the messenger RNA levels of MMP 9, VEGF-C, and VEGF-D were significantly suppressed by DcR3 knockdown by 90% when compared with the mock control (P < 0.05).
Loss of DcR3 impaired the growth and invasive property of HCC cell line of HepG2. Targeting DcR3 may be a potential therapeutic approach for the treatment of HCC.
诱饵受体3（DcR3）与Fas配体（FasL）结合并抑制FasL诱导的细胞凋亡。该受体在肝细胞癌（HCC）中过表达，且与肿瘤的生长和转移扩散相关。DcR3有望成为治疗HCC的新靶点，但关于DcR3致癌特性的分子机制知之甚少。因此，本研究探讨了DcR3在调节肝癌细胞HepG2生长和侵袭特性中的作用。
用基于慢病毒的靶向DcR3的短发夹RNA载体稳定转染HepG2细胞。在确认DcR3被敲低后，在体外评估细胞增殖、克隆形成、穿过Transwell膜的迁移能力和伤口愈合情况。还研究了DcR3敲低后的基质金属蛋白酶-9（MMP 9）、血管内皮生长因子（VEGF）-C和D的表达。多组间比较采用单因素方差分析（ANOVA），两两比较采用Student's t检验。P<0.05被认为具有统计学意义。
与其他肝癌细胞系和正常肝细胞Lo-2相比，DcR3在HepG2中过表达。与未敲低DcR3的细胞相比，DcR3的稳定敲低减缓了HepG2的生长（P<0.05），并使形成的克隆数减少了50%（P<0.05）。与对照相比，敲低还使HepG2穿过Transwell基质膜的迁移减少了五倍（P<0.05），并抑制了划痕伤口的闭合（P<0.05）。此外，与mock对照相比，DcR3敲低使MMP 9、VEGF-C和VEGF-D的信使RNA水平显著降低了90%（P<0.05）。
DcR3的缺失损害了HepG2肝癌细胞系的生长和侵袭特性。靶向DcR3可能是治疗HCC的一种潜在治疗方法。
