蝎毒多肽提取物体外抑制胰腺癌细胞侵袭转移及相关机制

目的研究蝎毒多肽提取物PESV对人高转移胰腺癌细胞MIA-PaCa2-3转移相关能力的影响及其作用的机制。为PESV在临床上治疗胰腺肿瘤提供理论根据。方法以高转移的人胰腺癌细胞系MIA-PaCa2-3为研究对象,通过细胞增殖实验(MTT法)观察在PESV影响下肿瘤细胞活性及增殖情况,通过细胞黏附实验、细胞划痕实验及Transwell体外侵袭实验检测PESV干预后细胞的黏附、侵袭及运动能力变化。免疫细胞化学实验及Western blot检测胰腺癌细胞的E-钙粘蛋白、纤粘蛋白和基质金属蛋白酶-9的表达。结果细胞增殖实验结果显示,PESV对胰腺癌细胞呈明显剂量及时间效应关系的增殖抑制作用,与对照组比较细胞增殖抑制作用明显(P<0.05);黏附实验表明PESV可降低胰腺癌细胞的黏附力,侵袭实验及划痕实验的结果表明PESV可以使细胞的侵袭力和运动能力下降,PESV(40mg.L-1)干预8h后,黏附力、侵袭力和运动能力的抑制率分别为(30.3±4.7)%、(42.1±3.7)%、(47.6±3.3)%,与对照组比较差异均有显著性(P<0.05);免疫细胞化学及Western blot实验检测均显示,PESV干预后,胰腺癌细胞E-钙粘蛋白阳性表达细胞数量上升,灰度值明显上调(P<0.05),FN及MMP-9蛋白阳性表达数量减少,灰度值明显下调(P<0.05)。结论PESV能够明显抑制胰腺癌细胞的增殖、黏附力、侵袭及运动能力,其机制可能是通过提高E-cad表达、降低FN和MMP-9表达而实现的。

The inhibitive effect of polypeptide extract from sorpion venom(PESV) on invasion and metastasis of pancreatic cancer cells in vitro and its primary mechanism

Aim To investigate the inhibitory effect and its possible mechanism of PESV on the metastasis-associated ability of human highly metastatic pancreatic carcinoma MIA-PaCa2-3 cells in vitro,and to provide the theoretical basis for clinical application.Methods MIA-PaCa2-3 pancreatic cancer cell line was chosen with highly metastasis ability as the object.The cell viability was determined by MTT assay and the state of cell proliferation after PESV interference was analyzed.Matrigel invasion assay,Wound Healing assay and Cell adhesion assay were used to detect the migration,the adhesion and the invasion abilities of these cells.To investigate the potential molecule mechanism,Immunocytochemistry-staining and Western blot were used to detect E-cadherin,fibronectin and matrix metalloproteinases-9 positive cells,after these cells were treated by the drugs.Results PESV could inhibit the proliferation of MIA-PaCa2-3 cells in a dose-time dependent manner.It also could inhibit the adhesion,invasion and migration capability.Their inhibitory ratio after treatment with 40 mg·L-1 PESV for 24 h was(30.3±4.7)%,(42.1±3.7)%,(47.6±3.3)% respectively.Significant changes in the activation of E-cad were detected by Immunocytochemistry and Western blot.The positive expressions of FN and MMP-9 proteinum were down regulated after the drug interference.Conclusions PESV can obviously suppress the pancreatic cancer cell proliferation.It can also inhibite the capability of migration,adhesion and invasion capability.The potential molecule mechanism is perhaps that PESV can enhance the E-cad expression,and reduce FN or MMP-9 expression.