Liu Xiaohua, Zhang Yincheng, Ren Wenhao, Cao Tengteng, Zhu Yongjin
Department of Oral and Maxillofacial Surgery, Xi'an Jiaotong University Stomatology Hospital, Xi'an 710004, Shaanxi Province, China.
J Biomed Res. 2010 May;24(3):215-22. doi: 10.1016/S1674-8301(10)60031-0.
To knockdown the C-erbB2 gene in salivary gland adenoid cystic carcinoma SACC-83 cells using RNA interference, and determine the effect of silencing C-erbB2 on cell proliferation.
C-erbB2-siRNA was transfected into SACC-83 cells. RT-PCR and immunohistochemistry were used to detect C-erbB2 expression in SACC-83 cells. Cell proliferation was measured by the MTT assay and gene knockdown was achieved by RNA interference. Apoptosis was analyzed by flow cytometry.
Compared with the control, C-erbB2 mRNA expression was decreased in the C-erbB2-siRNA transfection group, and immunohistochemical analysis indicated that C-erbB2 protein expression was decreased. After C-erbB2-siRNA was transfected for 48 h, absorbance at 570 nm (MTT) was 0.185±0.021 compared with 0.354±0.034, 0.299±0.053, and 0.314±0.049 in the blank control, liposome control and negative control siRNA groups, respectively. The differences were statistically significant (P < 0.05) between the C-erbB2-siRNA group and the control groups. Following the C-erbB2 knockdown, the percentage of apoptotic cells was 5.63% compared with 2.04%, 2.85%, and 2.98% in the three control groups, respectively. Proliferation of SACC-83 cells was inhibited, and early apoptotic cells were increased.
RNA interference can effectively silence C-erbB2 gene expression and inhibit growth of SACC-83 cells, which indicates the potential of targeting this gene as a novel gene therapy approach for the treatment of salivary gland adenoid cystic carcinoma.
利用RNA干扰技术敲低涎腺腺样囊性癌SACC-83细胞中的C-erbB2基因，并确定沉默C-erbB2对细胞增殖的影响。
将C-erbB2-siRNA转染至SACC-83细胞。采用RT-PCR和免疫组织化学方法检测SACC-83细胞中C-erbB2的表达。通过MTT法检测细胞增殖情况，利用RNA干扰实现基因敲低。采用流式细胞术分析细胞凋亡情况。
与对照组相比，C-erbB2-siRNA转染组中C-erbB2 mRNA表达降低，免疫组织化学分析表明C-erbB2蛋白表达降低。C-erbB2-siRNA转染48小时后，570nm处的吸光度（MTT）为0.185±0.021，而空白对照组、脂质体对照组和阴性对照siRNA组分别为0.354±0.034、0.299±0.053和0.314±0.049。C-erbB2-siRNA组与对照组之间差异具有统计学意义（P<0.05）。敲低C-erbB2后，凋亡细胞百分比为5.63%，而三个对照组分别为2.04%、2.85%和2.98%。SACC-83细胞的增殖受到抑制，早期凋亡细胞增加。
RNA干扰可有效沉默C-erbB2基因表达并抑制SACC-83细胞生长，这表明靶向该基因作为涎腺腺样囊性癌新的基因治疗方法具有潜力。
