橄榄苦苷调节IL-6/STAT3信号通路对肺癌细胞恶性生物学行为的影响

Effect of oleuropein on the malignant biological behavior of lung cancer cells by regulating IL-6/STAT3 signaling pathway

目的 初步探讨橄榄苦苷(Ole)调节白细胞介素-6(IL-6)/信号转导与转录激活子3(STAT3)信号通路对肺癌细胞恶性生物学行为的影响。方法 MTT法检测细胞增殖抑制率,筛选Ole最佳干预浓度;将A549细胞分为对照组、Ole低、中、高剂量组、rIL-6组(Ole高剂量+IL-6/STAT3通路激活剂)。A549细胞增殖、迁移、侵袭和凋亡依次用EdU染色、Transwell、流式细胞术检测;增殖细胞核抗原(PCNA)、基质金属蛋白酶-2(MMP-2)、切割型半胱氨酸天冬氨酸蛋白水解酶-3(Cleaved Caspase-3)及IL-6/STAT3信号通路蛋白表达用Western Blot法检测;构建肺癌移植瘤模型,测定移植瘤的体积和质量,免疫组化检测移植瘤中Ki-67、基质金属蛋白酶-9(MMP-9)蛋白表达,TUNEL染色检测肿瘤组织细胞凋亡。结果 与对照组比较,Ole低、中、高剂量组EdU阳性细胞率、迁移与侵袭细胞数、PCNA、MMP-2、IL-6、p-STAT3/STAT3表达显著降低,细胞凋亡率、Cleaved Caspase-3表达显著升高(P<0.05);IL-6/STAT3通路激活剂rIL-6减弱了Ole对A549恶性生物学行为抑制作用(P<0.05);体内实验表明,Ole显著降低移植瘤质量、体积及Ki-67、MMP-9、IL-6、p-STAT3/STAT3蛋白表达,升高肿瘤细胞凋亡率(P<0.05)。结论 Ole在体内体外可能通过抑制IL-6/STAT3信号通路,进而抑制A549细胞的恶性生物学行为。

Objective To investigate the effect of oleuropein(Ole)on the malignant biological behavior of lung cancer cells by regulating the interleukin-6(IL-6)/signal transducer and activator of transcription 3(STAT3)signaling pathway.Methods MTT method was applied to detect cell proliferation inhibition rate,and the optimal intervention concentration of Ole was screened.A549 cells were separated into the control group,the low,medium,and high Ole dose groups,and the rIL-6 group(Ole high dose+IL-6/STAT3 pathway activator).The proliferation,migration,invasion and apoptosis of A549 cells were detected by EdU staining,Transwell and flow cytometry,respectively.The protein expressions of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),cleaved caspase-3 and IL-6/STAT3 signaling pathway were detected by Western Blot.A lung cancer transplant tumor model was constructed,and the volume and mass of the transplanted tumor were measured.Immunohistochemical detection was applied to detect the expression of Ki-67 and matrix metalloproteinase-9(MMP-9)proteins in transplanted tumors.TUNEL staining was applied to detect apoptosis in tumor tissue cells.Results Compared with the control group,the EdU positive cell rate,migration and invasion cell counts,PCNA,MMP-2,IL-6,p-STAT3/STAT3 expression,and apoptosis rate,and the expression of Cleaved Caspase-3 in the low,medium,and high Ole dose groups were significantly lower(P<0.05).The IL-6/STAT3 pathway activator rIL-6 weakened the inhibitory effect of Ole on the malignant biological behavior of A549(P<0.05).In vivo experiments,it showed that Ole obviously reduced the mass and volume of transplanted tumors,and the expression of Ki-67,MMP-9,IL-6,p-STAT3/STAT3 proteins,and increased the apoptosis rate of tumor cells(P<0.05).Conclusion Ole can inhibit the malignant biological behavior of A549 cells by inhibiting the IL-6/STAT3 signaling pathway in vitro and in vivo.