摘要
目的:探讨STAT3及c-myc在肝癌细胞SMMC-7721中的表达,观察抗体干预对细胞增殖和凋亡的影响以及二者的相互关系。方法:免疫细胞化学及RT-PCR观察STAT3及c-myc在肝癌细胞中的表达;抗体干预后,MTT法检测细胞增殖,FCM检测细胞凋亡,RT-PCR检测STAT3及c-myc mRNA表达的变化。结果:STAT3及c-myc在肝癌细胞SMMC-7721中均有表达,单独应用STAT3或c-myc抗体以及联合应用2种抗体对细胞的增殖、凋亡均有影响,在48 h处理组,联合应用抗体的作用更明显。应用STAT3抗体作用于细胞,RT-PCR检测显示c-myc mRNA的表达下降,联合应用抗体后c-myc mRNA的表达明显降低。结论:肝癌细胞SMMC-7721中存在STAT3及c-myc信号通路,STAT3通过上调c-myc而抑制肝癌细胞的凋亡并促进其增殖,二者在肝癌的发生、发展中起重要作用。
Objective： To investigate the expressions of STA73 （ signal transducers and activators of transcription 3 ） and c-myc gene in hepatocellular carcinoma SMMC-7721 cells, observe the effects of antibody interference on cell proliferation and apoptosis, and analyze the correlation of the two genes. Methods： The protein and mRNA expressions of STAT3 and c-myc in SMMC-7721 cells were detected by immunocytochemical method and semi-quantitative RT-PCR, respectively. After antibody interference, cell proliferation was assessed by MTT assay ; apoptosis was detected by flow cytometry ; the mRNA expressions of STAT3 and c-myc were determined by RT- PCR. Results： Both STAT3 and c-myc were expressed in hepatocellular carcinoma SMMC-7721 cells. Immunocytochemical analysis showed that STAT3 and c-myc proteins were mainly localized in the cytoplasm. Proliferation and apoptosis of SMMC-7721 cells were inhibited by treatment with anti-STAT3 antibody and anti-c-myc antibody alone or in combination. The inhibitory effect was more significant for combined therapy group at 48 h. RT-PCR demonstrated that c-myc mRNA expression had significantly decreased after treatment with anti- STAT3 antibody alone and it further decreased after combined treatment. Conclusion：Our findings demonstrated that STAT3 and c-myc signaling pathway exists in SMMC-7721 cell line. STAT3 inhibites the apoptosis and facilitates the proliferation of SMMC-7721 cells by upregulating c-myc expression. STAT3 and c-myc play a critical role in the initiation and progression of hepatocellular carcinoma.
出处
《肿瘤》
CAS
CSCD
北大核心
2008年第5期394-397,共4页
Tumor
