胰腺星状细胞对胰腺癌AsPC-1细胞侵袭转移的影响
Effect of pancreatic stellate cells on invasion and metastasis of human pancreatic cancer cell line AsPC-1
摘要目的 探讨胰腺星状细胞(PSCs)对胰腺癌细胞侵袭转移的影响及基质细胞因子-I(SDF-1)在此过程中的作用.方法 常规分离、培养PSCs,收集及浓缩PSCs条件培养液(PSC-CM),应用不同浓度的PSC-CM、抗SDF-1抗体(anti-SDF-1)及两者联合应用处理AsPC-1细胞,采用MTT法检测AsPC-1细胞的增殖,Transwell小室检测细胞迁移,体外侵袭实验观察细胞侵袭能力.结果 对照组及0.25、0.5、1 μg/μl PSC-CM组AsPC-1细胞增殖的吸光度值(A490)分别为0.437 ±0.041、0.472±0.048、0.553±0.057、0.690±0.051,PSC-CM呈剂量依赖性促进细胞增殖,其中0.5、1μg/μl PSC-CM组与对照组间以及0.5 μg/μl与1μg/μl PSC-CM组间的差异具有统计学意义(P值均<0.05).对照组、anti-SDF-1组、PSC-CM组及PSC-CM± anti-SDF-1组细胞增殖的A490值分别为0.407±0.028、0.416±0.030、0.629±0.048、0.481±0.049;穿膜细胞数分别为(35.3±7.1)、(34.8±5.6)、(140.9±12.7)、(56.5±5.9)个;侵袭细胞数分别为(27.1±2.9)、(29.1±4.2)、(81.5±8.2)、(46.4±4.4)个.anti-SDF-1组与对照组间的差异无统计学意义,PSC-CM组细胞的增殖、迁移、侵袭能力较对照组显著增强(P<0.05或P<0.01),PSC-CM± anti-SDF-1组细胞的增殖、迁移、侵袭能力较PSC-CM组显著降低,但仍显著高于对照组(P<0.05或P<0.01).结论 PSC-CM可促进AsPC-1的增殖、迁移及侵袭,其机制为部分通过SDF-1/CXCR4受体配体系统.
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abstractsObjective To investigate the effect of pancreatic stellate cells on invasion and metastasis of human pancreatic cancer cell AsPC-1,and to determine the role of SDF-1 in this process.Methods PSCs were routinely isolated and cultured,and PSCs conditioned media(PSC-CM) was collected and concentrated.Different concentrations of PSC-CM,anti SDF-1 and their combination were used to treat AsPC-1 cells,and MTT assay was applied to detect the proliferation of pancreatic cancer cells.Transwell chamber migration assay was employed to detect the migration of AsPC-1 cells.In vitro invasion assay was used to determine the invasion of AsPC-1 cells.Results A490 values of AsPC-1 cell in control group and 0.25,0.5,1 μg/lμl PSCCM group were 0.437 ±0.041,0.472 ±0.048,0.553 ±0.057,0.690 ±0.051,and PSC-CM promoted cell proliferation in a dose dependant manner.The difference between 0.5,1 μg/μl PSC-CM group and control group,and between 1 μg/l and 0.5 μg/μl PSC-CM group was statistically significant (P<0.05).A490 values of control group,anti SDF-1 group,PSC-CM group and PSC-CM ± anti SDF-1 group were 0.407 ±0.028,0.416 ±0.030,0.629 ±0.048,0.481 ±0.049.The numbers of penetrating cells were 35.3 ±7.1,34.8±5.6,140.9 ± 12.7,56.5±5.9,and the numbers of invasive cells were 27.1 ±2.9,29.1 ±4.2,81.5 ±8.2,46.4 ± 4.4.The difference between anti SDF-1 group and control group was not statistically significant.The proliferation,migration and invasion of pancreatic cancer cells in PSC-CM group was significantly higher than those of control group (P <0.05 or P <0.01).The proliferation,migration and invasion of pancreatic cancer cells in PSC-CM ± anti SDF-1 group was significantly lower than those of PSC-CM group,but they were significantly higher than those of control group (P < 0.01).Conclusions PSCs can promote proliferation,migration and invasion of pancreatic cancer cells AsPC-1,and the mechanism may be partly due to SDF-1/CXCR4 receptor ligand axis.
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