Adenovirus-mediated RhoA shRNA suppresses growth of esophageal squamous cell carcinoma cells in vitro and in vivo.

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.