Tomizawa Minoru, Shinozaki Fuminobu, Sugiyama Takao, Yamamoto Shigenori, Sueishi Makoto, Yoshida Takanobu
Departments of Gastroenterology.
Exp Ther Med. 2010 Sep;1(5):863-866. doi: 10.3892/etm.2010.131. Epub 2010 Jul 21.
To suppress the invasion of hepatocellular carcinoma (HCC) cells into surrounding connective tissues during metastasis, we investigated the usefulness of sorafenib. In order to search for model cell lines, cell numbers were counted to reveal cell lines with the potential to proliferate in serum-free media. Cell proliferation and cell motility were analyzed with the MTS and wound assay, respectively. 5-Bromo-2'-deoxyuridine (BrdU) labeling and mitotic and apoptotic indices were analyzed to assess the cell cycle and apoptosis. The expression levels of cyclin D1 and the cleavage of caspase-3 were analyzed by Western blotting. HLF cells exhibited growth in the serum-free medium, while the other cell lines examined did not. Sorafenib suppressed the cell proliferation and motility of the HLF cells in the serum-free media. Both indices of BrdU and mitotic potential decreased and the apoptotic index was increased in the serum-free media with sorafenib, suggesting that the cell cycle was suppressed and apoptosis was induced. The expression levels of cyclin D1 decreased and the cleavage of caspase-3 was noted in the serum-free media with sorafenib. Sorafenib may be suitable for molecular therapy to suppress the metastasis of HCC.
为了抑制肝细胞癌(HCC)细胞在转移过程中侵入周围结缔组织,我们研究了索拉非尼的有效性。为了寻找模型细胞系,通过计数细胞数量来揭示有潜力在无血清培养基中增殖的细胞系。分别用MTS法和划痕试验分析细胞增殖和细胞运动能力。通过5-溴-2'-脱氧尿苷(BrdU)标记以及有丝分裂和凋亡指数分析来评估细胞周期和凋亡情况。通过蛋白质印迹法分析细胞周期蛋白D1的表达水平和半胱天冬酶-3的切割情况。HLF细胞在无血清培养基中表现出生长,而所检测的其他细胞系则没有。索拉非尼在无血清培养基中抑制了HLF细胞的增殖和运动能力。在含有索拉非尼的无血清培养基中,BrdU指数和有丝分裂潜能均降低,凋亡指数增加,这表明细胞周期受到抑制且诱导了凋亡。在含有索拉非尼的无血清培养基中,细胞周期蛋白D1的表达水平降低且观察到半胱天冬酶-3的切割。索拉非尼可能适用于抑制HCC转移的分子治疗。