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- 2016
沉默肝星状细胞神经菌毛素-1抑制肝癌生长的实验研究
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Abstract:
摘要：目的 探讨沉默肝星状细胞神经菌毛素-1(neuropilin-1, NRP-1)表达后，是否会影响其对肝癌的促生长作用，并初步探讨可能存在的作用机制。方法 细胞免疫荧光及细胞荧光双重染色观察NRP-1在肝星状细胞LX2中的表达及其与血小板源性生长因子受体β(platelet-derived growth factor receptor-β, PDGFR-β)的共表达；MTT法检测沉默肝星状细胞NRP-1后对肝癌细胞体外增殖能力影响；建立裸鼠皮下肝癌移植瘤模型，绘制生长曲线，免疫组化法观察各组瘤体α-平滑肌肌动蛋白(α-smooth muscle actin, α-SMA)、NRP-1、PDGFR-β表达强度的差异。结果 LX2表面存在NRP-1表达，且NRP-1与PDGFR-β共表达于LX2；沉默LX2 NRP-1后对HepG2促增殖作用降低(P<0.05)；NRP-1 KG移植瘤体积较NRP-1 CG、NRP-1 NG小(P<0.05)，进一步对各组瘤体行免疫组化染色并进行表达强度积分分析，发现NRP-1 KG瘤体间质NRP-1、PDGFR-β表达均较NRP-1 CG弱(χ2=25.89，P<0.05；χ2=28.12，P<0.05)。结论 沉默肝星状细胞NRP-1表达后，其对肝癌细胞促增殖作用降低，在体内环境中其对肝癌皮下移植瘤促生长作用也降低，其机制与肝星状细胞活化减弱有关，而肝星状细胞NRP-1表面共受体PDGFR-β表达减弱也可能参与这一过程。
ABSTRACT: Objective To investigate whether the expression of hepatic stellate cells neuropilin-1 silenced (NRP-1) affects the growth of hepatocellular carcinoma cells (HCCs) and to explore the possible mechanism. Methods Expression of NRP-1 and co-expression with platelet-derived growth factor receptor-β(PDGFR-β) in hepatic stellate cells were observed by immunofluorescence and fluorescent double staining. The effect of silencing NRP-1 of HSCs on the proliferation of HCC cells was detected by MTT in vitro. A xenograft hepatocellular carcinoma model of nude mouse was established subcutaneously. The growth curve was drawn and the expressions of α-SMA, NRP-1 and PDGFR-β in tumors were observed by immunohistochemistry staining after the mice were killed. Results NRP-1 was expressed in the membrane of LX2 and was co-expressed with PDGFR-β. Silencing NRP-1 of LX2 reduced the proliferation of HepG2 in vitro (P<0.05). The tumor volume in NRP-1 KG group reduced obviously compared with that in NRP-1 CG group (P<0.05). The expressions of NRP-1 and PDGFR-β in NRP-1 KG group were weaker than those in NRP-1 CG group by immunohistochemistry (χ2=25.89, P<0.05; χ2=28.12, P<0.05). Conclusion Silencing NRP-1 expression of HSCs can decrease its effect on the proliferation of HCCs in vitro and the growth-promoting effect of HSCs on HCCs is also decreased in vivo, which is due to the inhibition of HSCs activation, and the decreased expression of co-receptor PDGFR-β may play a role in this process
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