p75NGFR对胰腺癌细胞株SW1990生物学行为的影响

Biological effects of p75NGFR on pancreatic cancer cell line SW1990

目的通过体内和体外实验,探讨p75NGFR在胰腺癌SW1990细胞生长、分化以及侵袭中的作用。方法采用脂质体转染法将含p75NGFR的真核表达质粒转染人胰腺癌细胞株SW1990,建立稳定表达p75NGFR的细胞模型;应用MTT法、平板克隆形成实验以及流式细胞技术分别检测细胞的生长曲线、克隆形成能力以及细胞周期变化;建立胰腺癌的裸鼠皮下移植瘤和胰腺原位移植瘤模型,观察p75NGFR对SW1990细胞裸鼠成瘤能力、肿瘤组织学分化、细胞侵袭和转移的影响。结果与对照组相比较,转染p75NGFR的SW1990细胞其生长速度缓慢(P<0.01),克隆形成能力降低(P<0.05);转染细胞的细胞周期改变表现为G1期细胞明显增多和S期细胞明显减少(均为P<0.01)。与对照组相比较,转染组裸鼠皮下以及胰腺原位的成瘤率以及肿瘤体积减小(P<0.01),肿瘤的生长缓慢,组织学分化差,浸润性生长趋势不明显,未见肿瘤卫星现象。结论体外实验均发现p75NGFR通过阻滞细胞周期进程而显著抑制胰腺癌细胞株的生长,体内研究发现p75NGFR可抑制裸鼠皮下及胰腺原位移植瘤的形成、分化和侵袭,提示p75NGFR与胰腺癌的生物学行为密切相关。

Purpose To evaluate the biological effects of p75NGFR on the growth, differentiation and invasion of SW1990 cell line in vitro and in vivo. Methods A stably-transfected cell line of SW1990, SW1990/p75NGFR, which could express p75NGFR, was established using the Lipefectamine method. Growth curve, clony formation and flow cytometry were performed to investigate the growth ability and cell cycle of transfected cells. Subcutaneous and intrapancreatic allograft models in nude mice were established to investigate the effects of p75NGFR on tumorigenesis, tumor growth, invasion and metastasis of SW1990 in vivo. Results Compared with control groups, SW1990/p75NGFR showed slower growth pattem （P 〈0. 01 ） and poorer ability of colony formation （P 〈0. 01 ）. Flow cytometry results revealed that the exogenous p75NGFR could significantly accumulate cells in G0/G1 stage and decrease cell amounts in S phase （P〈0. 01）. In vivo, SW1990/p75NGFR also owned less capability of tumorogenesis both in subcutaneous tissue and in the pancreas. Tumor size was significantly Smaller and the incidence of intrapancreatic allografts formation remarkably decreased （ P = 0. 00） while compared with controls. And no ascite, satellite lesions, vascular invasion and nearby normal pancreatic tissue invasion were found in the tumors induced by SW1990/p75NGFR transfection. Conclusions p75NGFR could strikingly inhibit the growth of SW1990 cell line mainly by cell cycle arrest in vitro and impact the formation, differentiation and invasion of subcutaneous and intrapancreatic allografts of SW1990 cell in vivo. These findings indicate that p75NGFR may play a very important role in the biological behavior of pancreatic adenocarcinoma.