中文摘要：

目的探讨针对人宫颈癌基因（HCCR）反义核苷酸影响肝癌细胞增殖、凋亡的作用。方法构建pcD—NA3．1-HCCR反义真核表达质粒。转染HepG2细胞，实时定量PCR检测HCCR mRNA表达水平，Western blot检测其蛋白表达水平，流式细胞仪观察HepG2细胞凋亡和细胞周期的变化．MTT法检测细胞的增殖活性。结果HCCR反义核酸可有效抑制HepG2细胞HCCR的表达，与转染前相比，转染后24小时HCCR mRNA水平下降到16％，其蛋白表达水平在24小时也明显降低。对细胞的增殖与凋亡活性检测显示：转染HCCR反义质粒后，HepG2细胞增殖受到抑制．转染后24小时抑制率为47．62%（P〈0．05），细胞凋亡率为14．34％±0．91％．较对照组明显增多（t=21．799，P〈0．05），细胞分裂多停止在G0G1期。结论HCCR反义核酸可以抑制HepG2细胞中HCCR的表达．抑制细胞增殖，促进细胞凋亡，使细胞停滞于G1期。HCCR反义核酸用于肝癌细胞的基因治疗具有一定的潜在意义。

英文摘要：

Objective To investigate the effect of antisense nucleotide of human cervical cancer oncogene （HCCR） on the cell proliferation and apoptosis of HepG2 cells. Methods Antisense expression vector of pCDNA3. 1-HCCR was constructed and transfected into HepG2 cells. The expression level of HCCR mRNA and protein were detected by real time PCR and Western blot. The effect of the pCDNA3. 1-HCCR on cell proliferation was assayed by MTT method and the cell apoptosis was observed by flow cytometry. Results Real time PCR and Western blot analysis showed that antisense nucleotide of HCCR could effectively suppress HCCR expression level compared with those of non-transfected and the HCCR mRNA level at 24h after transfection was reduced to 16%, and the level of HCCR protein was also decreased . MTT method confirmed the cell proliferation of HepG2 cells were inhibited（P〈0.05）, with the inhibition rate at 24h after transfection of 47. 6%, while the apoptosis by flow cytometry was 1.74%±0. 44%, 2. 10%±0. 30%, 14. 34%±0.91 % in HepG2 cells group, HepG2/pCDNA3.1（ ＋ ） group and pCDNA3.1 （＋）-HCCR group, respectively , and the apoptosis of HepG2 cells transfeeted with pCDNA3. 1（＋ ）-HCCR increased much more than control groups（t= 21. 799, P 〈0.05）. Cell-cycle analysis by flow cytometry showed that transfected HepG2 cells were arrested at the G1 phase of the cell cycle. Condusion The antisense nucleotide of HCCR can effectively inhibit the expression level of HCCR mRNA and protein , suppress cell proliferation and induce cell apoptosis and antisense nucleotide of HCCR may serve as a potential antitumor strategy in HCCs.