Hsieh Yi-Hsien, Wu Trang-Tiau, Huang Chih-Yang, Hsieh Yih-Shou, Liu Jer-Yuh
Institute of Biochemistry and Biotechnology, Chung-Shan Medical University, Taichung, Taiwan, ROC.
Chin J Physiol. 2007 Feb 28;50(1):9-15.
Our previous studies have found that reducing expression of myeloid zinc finger-1 (MZF-1) inhibited protein kinase C alpha (PKCalpha) expression and decreased cell migration and invasion in human hepatocellular carcinoma (HCC). In this study, we further investigated the role of MZF-1 in tumorigenesis. The SK-Hep-1 HCC cells were transfected with the antisense oligonucleotide (ODN) of MZF-1. The pretreated cells was then detected the mRNA and protein levels by RT-PCR and western blotting, and the cell growth was assayed by MTT. Meanwhile, the pretreated-SK-Hep-1 HCC cells were implanted subcutaneously into nude mice to observe the tumor growth and calculated tumor inhibitory rate. The results showed that, at the dosage of 5 microM, the antisense ODN MZF-1 suppressed both MZF-1 and PKCalpha productions by SK-Hep-1 HCC cells after cationic liposome-mediated transfection, to 15% and 30% in control cultures of 0 microM dosage, respectively. The growth of SK-Hep-1 HCC cells was inhibited by the 2-5 microM doses of the antisense ODN MZF-1, whereas the control reagent, the sense ODN MZF-1, showed no effects. In BALB/nude mice, SK-Hep-1 HCC cells pretreated with the 5 microM antisense ODN MZF-1 formed tumors much smaller than the cells with sense ODN. The mean of inhibitory rate of tumor growth was 71.2 +/- 8.6%, and the tumor formation time was prolonged from 14 days to 26 days. These findings suggested the usefulness of antisense ODN MZF-1 as a new reagent for cancer therapy.
我们之前的研究发现，降低髓样锌指蛋白-1（MZF-1）的表达可抑制蛋白激酶Cα（PKCα）的表达，并减少人肝细胞癌（HCC）中的细胞迁移和侵袭。在本研究中，我们进一步探讨了MZF-1在肿瘤发生中的作用。用MZF-1的反义寡核苷酸（ODN）转染SK-Hep-1肝癌细胞。然后通过RT-PCR和蛋白质印迹法检测预处理细胞的mRNA和蛋白质水平，并通过MTT法测定细胞生长情况。同时，将预处理后的SK-Hep-1肝癌细胞皮下植入裸鼠体内，观察肿瘤生长情况并计算肿瘤抑制率。结果显示，在5微摩尔剂量下，阳离子脂质体介导转染后，反义ODN MZF-1可将SK-Hep-1肝癌细胞中MZF-1和PKCα的产生分别抑制至0微摩尔剂量对照培养物中的15%和30%。2-5微摩尔剂量的反义ODN MZF-1可抑制SK-Hep-1肝癌细胞的生长，而对照试剂正义ODN MZF-1则无此作用。在BALB/n裸鼠中，用5微摩尔反义ODN MZF-1预处理的SK-Hep-1肝癌细胞形成的肿瘤比用正义ODN处理的细胞形成的肿瘤小得多。肿瘤生长抑制率的平均值为71.2±8.6%，肿瘤形成时间从14天延长至26天。这些发现表明反义ODN MZF-1作为一种新型癌症治疗试剂具有应用价值。
