摘要
目的 探讨锌指和BTB结构域蛋白12(ZBTB12)对肝细胞癌(HCC)细胞有氧糖酵解、增殖和迁移的影响。方法 利用实时荧光定量聚合酶链反应(qPCR)、Western blotting和免疫组化检测ZBTB12在HCC组织和配对癌旁组织中的表达差异。构建敲低ZBTB12的HCC细胞Hep3B和SMMC-7721,体外通过平板克隆形成实验、CCK-8实验和Transwell小室实验检测ZBTB12对HCC细胞增殖和迁移的影响。体内通过裸鼠皮下瘤模型和肺转移模型评估ZBTB12对HCC细胞增殖和迁移的影响。通过检测葡萄糖摄取量、乳酸产生量、耗氧率(OCR)和细胞外酸化率(ECAR)明确ZBTB12对HCC细胞有氧糖酵解的影响,进一步采用qPCR和Western blotting检测ZBTB12对糖酵解关键酶的影响。结果 ZBTB12的mRNA和蛋白水平在HCC组织中较癌旁组织均呈现明显的高表达。敲低ZBTB12明显抑制了Hep3B和SMMC-7721细胞在体外和体内的增殖和迁移的能力。机制研究表明敲低ZBTB12明显抑制了HCC细胞的葡萄糖摄取量、乳酸产生量、OCR和细胞外酸ECAR,进一步分析发现敲低ZBTB12明显抑制了糖酵解关键酶HK2和LDHA的表达。结论 ZBTB12的表达在HCC组织中明显上调,敲低ZBTB12可以明显抑制HCC细胞的有氧糖酵解、增殖和迁移,有望成为HCC治疗的潜在靶点。
Objective To investigate the effects of zinc finger and BTB domain containing 12(ZBTB12) on aerobic glycolysis, proliferation and migration of hepatocellular carcinoma(HCC) cells. Methods Quantitative real-time polymerase chain reaction(qPCR), Western blotting and immunohistochemistry were used to detect the expression difference of ZBTB12 in paired human HCC and paratumor tissues. Human HCC cell lines Hep3B and SMMC-7721 were chosen to construct ZBTB12 knockdown cell lines. Effects of ZBTB12 on the proliferation and migration of HCC cells were detected by colony formation assay, CCK-8 assay and Transwell assay in vitro, as well as the subcutaneous tumor model and lung metastasis model in nude mice in vivo. Finally, effects of ZBTB12 on aerobic glycolysis of HCC cells were determined by measuring glucose intake, lactic acid production, oxygen consumption rate(OCR) and extracellular acidification rate(ECAR). Influences of ZBTB12 on key glycolytic enzymes were detected by qPCR and Western blotting. Results ZBTB12 was upregulated in human HCC tissues compared with paratumor tissues. ZBTB12 knockdown significantly inhibited the aerobic glycolysis, proliferation and migration of Hep3B and SMMC-7721 cells. Further studies showed that ZBTB12 knockdown significantly suppressed the expressions of key glycolytic enzymes HK2 and LDHA. Conclusion ZBTB12 expression is significantly up-regulated in HCC tissues. Knocking down ZBTB12 can significantly inhibit the aerobic glycolysis, proliferation and migration of HCC cells, which is expected to become a potential target for HCC treatment.
作者
吴洁
吴群
高春
吴鑫阳
李国春
WU Jie;WU Qun;GAO Chun;WU Xinyang;LI Guochun(Department of Geriatrics,Nanjing Central Hospital,Nanjing 210009,China)
出处
《临床肿瘤学杂志》
CAS
2022年第12期1078-1084,共7页
Chinese Clinical Oncology
关键词
肝细胞癌
锌指和BTB结构域蛋白12
有氧糖酵解
增殖迁移
Hepatocellular carcinoma
Zinc finger and BTB domain containing 12
Aerobic glycolysis
Proliferation and migration
