摘要
目的探讨咪喹莫特(IMQ)对脑胶质细胞瘤U87细胞增殖的影响。方法U87细胞分为对照组、1 mmol/L IMQ组、5 mmol/L IMQ组、1 mmol/L IMQ+STAT3抑制剂(inhibitor)(STAT3-IN)组和5 mmol/L IMQ+STAT3-IN组。5-乙炔基-2’-脱氧尿苷(EdU)和MTT实验检测各组U87细胞EdU标记的细胞数目或增殖吸光度值;Real-time PCR、ELISA检测各组U87细胞白细胞介素6(IL-6)mRNA、肿瘤坏死因子-α(TNF-α)mRNA及其蛋白含量;Western blotting检测各组U87细胞STAT3、磷酸化STAT3(p-STAT3)、核因子(NF)-κB、磷酸化NF-κB(p-NF-κB)蛋白表达。结果相对于对照组,1 mmol/L IMQ组和5 mmol/L IMQ组U87细胞的EdU标记细胞数目和吸光度值依次降低,并呈现剂量依赖性(P<0.01,n=10),IMQ+STAT3-IN组和5 mmol/L IMQ+STAT3-IN组EdU标记的U87细胞数目、吸光度值均降低。1 mmol/L IMQ组和5 mmol/L IMQ组U87细胞STAT3、p-STAT3、NF-κB、p-NF-κB或IL-6、TNF-α蛋白均比对照组低表达(P<0.01,n=10),1mmol/L IMQ+STAT3-IN组和5 mmol/L IMQ+STAT3-IN组上述蛋白持续低表达(P<0.01,n=10)。结论咪喹莫特通过下调STAT3/NF-κB通路降低IL-6、TNF-α含量,进而抑制U87细胞增殖。
Objective To explore the effect of imiquimod(IMQ)on the proliferation of glioma U87 cell line U87.Methods U87 cells were divided into control group,1 mmol/L I mmol/L MQ group,5 mmol/L IMQ group,1 mmol/L IMQ+STAT3 inhibitor(STAT3-IN)group and 5 mmol/L IMQ+STAT3-IN group.To detect the number of 5-ethynyl-2’-deoxyuridine(EdU)-labeled cells or proliferation absorbance(A)values in each group by EdU and MTT assays.Interleukin(IL)-6 mRNA,tumor necrosis factor(TNF)-αmRNA and protein content in U87 cells of each group were detected by Real-time PCR or ELISA.Western blotting was used to detect the protein expression of STAT3,phosphorylated STAT3(p-STAT3),nuclear factor(NF)-κB and phosphorylated NF-κB(p-NF-κB)in U87 cells of each group.Results Compared with the control group,the number of EdU-labeled cells and absorbance values of U87 cells were successively decreased in 1 mmol/L IMQ group and 5 mmol/L IMQ group,showing a dose-dependent manner(P<0.01,n=10).However,the number of EdU-labeled cells and the A values in IMQ+STAT3-IN group and 5 mmol/L IMQ+STAT3-IN group were significantly reduced.Compared with the control group,the protein expression of STAT3,p-STAT3,NF-κB,p-NF-κB,IL-6 and TNF-αwere continuing low level in U87 cells of 1 mmol/L IMQ group and 5 mmol/L IMQ group(P<0.01,n=10).As well as in 1 mmol/L IMQ+STAT3-INgroup and 5 mmol/L IMQ+STAT3-IN group,the proteins of above were low expressed(P<0.01,n=10).Conclusion Imiquimod decreased the contents of IL-6 and TNF-αby down-regulating STAT3/NF-κB pathway,and thus inhibited the proliferation of U87 cells.
作者
王澍
尹璐
刘宏斌
徐加志
赵吉波
潘云志
孙玉荣
WANG Shu;YIN Lu;LIU Hong-bin;XU Jia-zhi;ZHAO Ji-bo;PAN Yun-zhi;SUN Yu-rong(Department of Neurology,the Third Affiliated Hospital of Qiqihar Medical College,Heilongjiang Qiqihar 161006,China;School of Pathology,Qiqihar Medical College,Heilongjiang Qiqihar 161006,China)
出处
《解剖学报》
CAS
CSCD
北大核心
2022年第3期323-329,共7页
Acta Anatomica Sinica
基金
齐齐哈尔医学科学院临床科研基金(QMSI2019L-29)。
