Inhibitory effect and mechanism of midazolam on transplanted tumor of nude mice with human lung cancer A549 cells
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摘要: 目的 探讨咪达唑仑对人肺癌A549细胞裸鼠移植瘤的抑制效应及机制。 方法 建立人肺癌A549细胞裸鼠移植瘤模型,共建模成功12只,分为咪达唑仑组与对照组各6只。咪达唑仑组腹腔注射咪达唑仑0.8 mg/kg, 1次/d; 对照组给予生理盐水注射。20 d后观察2组荷瘤裸鼠体质量和皮下移植瘤体积变化。处死裸鼠,运用苏木精-伊红染色法(HE)观察咪达唑仑对肺癌移植瘤细胞形态的作用效应; 依据细胞凋亡检测法(TUNEL)检测移植瘤中细胞凋亡情况; 采用免疫组织化学染色法(IHC)观察移植瘤组织细胞中核蛋白质Ki-67的改变,采用蛋白质印迹法(Western-blot)观察移植瘤组织中信号传导转录激活因子3(STAT3)的改变。 结果 与对照组相比,咪达唑仑组移植瘤质量、肿瘤体积均显著较小(P<0.05或P<0.01); HE染色显示咪达唑仑组移植瘤组织发生大面积坏死; TUNEL法显示咪达唑仑组细胞凋亡数显著高于对照组,IHC染色表明咪达唑仑干预后显著下调移植瘤组织中核蛋白质Ki-67蛋白的表达,咪达唑仑组STAT3蛋白表达显著下调(P<0.05)。 结论 咪达唑仑可能是通过调控STAT3的表达显著抑制A549移植瘤组织的生长,诱导肿瘤细胞凋亡。Abstract: Objective To explore the inhibitory effect and mechanism of midazolam on transplanted tumor of nude mice with human lung cancer A549 cells. Methods Totally 12 nude mice model of human lung cancer A549 cells were established and randomly divided into midazolam group(n=6)and control group(n=6). Dosage of 0.8 mg/kg midazolam was injected intraperitoneally once a day in midazolam group, and saline was injected in control group. After 20 days, the body mass and the volume of transplanted tumor were observed. Nude mice were sacrificed, and HE staining was used to observe the effect of midazolam on the cell morphology of transplanted tumor tissue. TUNEL staining was used to detect tumor cell apoptosis in transplanted tumor tissues, immunohistochemical(IHC)staining was used to observe the changes in Ki-67 in tumor tissue cells, and Western-blot detection was used to observe the changes of signal transducer and activator of transcription 3(STAT3)proteins in transplanted tumor tissue. Results Compared with the control group, the tumor mass and volume in midazolam group were significantly smaller(P<0.05 or P<0.01). HE staining showed that the tumor tissue of midazolam group had a large area of necrosis. TUNEL staining showed that the apoptosis number of midazolam group was significantly higher than that of the control group, IHC staining showed that midazolam can significantly reduced the expression of Ki-67 protein in the tumor tissue after intervention, midazolam can significantly reduced the expression of - Ki-67 protein in the tumor tissue, and the expression of STAT3 protein in midazolam group regulated down significantly(P<0.05). Conclusion Midazolam can significantly inhibit the growth of A549 transplanted tumor tissue and induce tumor cell apoptosis by regulating STAT3 expression.
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