Adenovirus-mediated RhoA shRNA suppresses growth of esophageal squamous cell carcinoma cells in vitro and in vivo
Adenovirus-mediated RhoA shRNA suppresses growth of esophageal squamous cell carcinoma cells in vitro and in vivo
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DOI：
10.1007/s12032-010-9774-y
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发表时间：
2012-03-01
期刊：
影响因子：
3.4
通讯作者：
Liu, Wenchao
中科院分区：
文献类型：
--
作者：
Ma, Ji;Zhang, Jian;Liu, Wenchao
RhoA在食管鳞癌中的过度表达提示预后不良，并与肿瘤-淋巴结-转移(TNM)临床分型有关。然而，到目前为止，RhoA在食管癌发生发展中的作用仍有待确定。我们使用腺病毒介导的针对人RhoA的小发夹RNA(ShRNA)(Ad-sh-RhoA)在蛋白质和mRNA水平上有效地沉默了RhoA表达的ECA-109 ESCC细胞中的靶基因表达。结果表明，Ad-sh-RhoA分别抑制了Eca-109细胞的增殖和迁移，并抑制了细胞的愈合。流式细胞仪检测，Ad-sh-RhoA可促进Eca-109细胞的凋亡，抑制细胞周期于G1-S期。最后，在裸鼠模型中，瘤内注射腺病毒携带的RhoA shRNA，每3天一次，连续20天，显著抑制了移植瘤Eca-109的生长和血管生成。综上所述，这些数据表明RhoA可能是ESCC细胞中的一个关键分子，因此，用腺病毒携带的shRNA特异性地抑制Rho信号通路是治疗侵袭性ESCC的一种有前途的方法。
Over-expression of RhoA in esophageal squamous cell carcinoma (ESCC) indicates a poor prognosis and is correlated with the tumor-node-metastasis (TNM) clinical classification. However, until now RhoA function in the ESCC progression remains to be established. We employed adenovirus-mediated small hairpin RNA (shRNA) against human RhoA (Ad-sh-RhoA) to efficiently silence target gene expression in RhoA-expressing Eca-109 ESCC cells at both protein and mRNA levels. Consequently, Ad-sh-RhoA reduced the proliferation and migration of Eca-109 cells assayed by MTT assay and cell wound healing, respectively. Moreover, Ad-sh-RhoA increased cell apoptosis and inhibited the cell cycle G1-S-phase progression of Eca-109 cells assessed by flow cytometry. Finally, in a nude mouse model, intratumoral injections of adenovirus-delivered RhoA shRNA every 3 days for 20 days significantly inhibited the growth and angiogenesis of xenografted Eca-109 tumors. In summary, these data indicate that RhoA may be a key molecule in ESCC cells, and thus, specific inhibition of the Rho signaling pathway with adenovirus-delivered shRNA represents a promising approach for the treatment of aggressive ESCC.