CD40调节胰腺癌细胞生物学行为的研究

Effects of CD40 pathway on the biological behavior of pancreatic cancer cells

目的:研究CD40L对胰腺癌细胞生物学特性、干性的影响。方法:CCK-8活细胞计数法分析CD40可溶性配体(s CD40L)不同浓度(0、1、2、4μg/m L)在不同时间对人胰腺癌细胞系panc02生长增殖的影响,Annexin V/PI双染法检测凋亡;细胞划痕实验检测迁移的改变。Q-PCR检测4μg/m L s CD40L处理panc02 48 h后c-Myc、OCT-4、Sox-2水平的改变。建立Balb/c裸鼠胰腺癌荷瘤模型,随机分为2组,对照组(生理盐水)和s CD40L(10 mg/kg)组,分别腹腔注射s CD40L 10mg/kg以及等体积生理盐水,每天注射3次,游标卡尺测量0、7、14、21、28 d时肿瘤体积。结果:与对照组比较,浓度为1、2、4μg/m L的s CD40L处理panc02 96 h时可显著抑制panc02的增殖(P<0.01),并促进panc02的凋亡(P<0.01),并呈剂量依赖性地抑制panc02的迁移能力(P<0.01);s CD40L 4μg/m L组可明显抑制panc02细胞的cMyc、OCT-4、Sox-2的表达(P<0.01)。在裸鼠荷瘤实验中,与对照组比较,s CD40L处理28 d明显抑制肿瘤体积(P<0.01,252±13 vs.189±9)。结论:CD40通路的活化能够抑制胰腺癌细胞增殖迁移、促进凋亡,同时抑制胰腺癌细胞系体内的增殖能力,而这一效应可能与CD40L抑制胰腺癌细胞系干性相关。

AIM：To study the effect of CD40 L on the biological characteristics and stenness of pancreatic cancer cells.METHODS：The effect of different concentrations of s CD40 L（0,1,2,4 μg/m L） on the proliferation of human pancreatic cancer cell line panc02 was analyzed by CCK-8 live cell counting assay.Apoptosis was detected by Annexin V/PI double staining;migrational ability was detected by Cell Scratch Assays.Q-PCR was used to detect the levels of c-Myc,OCT-4 and Sox-2 after panc02 treatment for 4 h/m L s CD40 L.A Balb/c nude mouse model of pancreatic cancer was established and randomly divided into two groups：control group（normal saline） and s CD40 L（10 mg/kg）group.They were injected intraperitoneally with s CD40 L 10 mg/kg and an equal volume of saline for three times a day.Tumor volume was measured using vernier calipers at 0,7 th,14 th,21 st,and28 th day.RESULTS：Compared with the control group,the growth and migration of panc02 decreased significantly after s CD40 L treatment（1,2,4 μg/m L for 96 h）,and apoptosis was promoted in a dose-dependent manner（P〈0.01）.s CD40 L（4μg/m L） significantly inhibited the expression of cMyc,OCT-4,Sox-2 of panc02（P〈0.01）.In the nude mice tumor-bearing experiment,compared with the control group,s CD40 L significantly inhibited the tumor volume at 28 th day（P〈0.01,252 ± 13 vs.189 ± 9）.CONCLUSION：Activation of CD40 pathway inhibits proliferation,migration and apoptosis of pancreatic cancer cells as well as the proliferation of pancreatic cancer cell lines in vivo.This effect may be related to the change of stemness in pancreatic cancer cell lines.