Objective: As the second common malignant tumor in females, cervical cancer severely affects public health. Epithelial-Mesenchymal Transition (EMT) occurs in tumor cells and is a key factor for tumor infiltration and metastasis. MicroRNA (miRNA) can inhibit mRNA translation and thus may be associated with tumor EMT. This study treated cervical cancer Hela cell line with neo-gambogic acid, to investigate its regulation on mRNA, aiming to elucidate its effects on EMT occurrence and drug resistance.
Materials and Methods: Human cervical carcinoma Hela cells were transfected with miRNA106b and miRNA93 vectors, and treated with different concentrations of neo-gambogic acid (0.5 μg/ml, 1.0 μg/ml, 1.5 μg/ml and 2.0 μg/ml) for different time points (24 h, 48 h and 72 h), followed by cisplatin treatment. Real-time PCR was used to detect the expression of miRNA. Cell invasion and migration were detected by Transwell assay, while Western blot was used to quantify the EMT-related protein expression.
Results: With higher concentration and/or longer treatment time of neo-gambogic acid, expressions of miRNA106b and miRNA93 were decreased (p<0.05) in Hela cells, which also had reduced cell invasion and migration ability (p<0.05). The higher concentration of neo-gambogic acid decreased the protein expressions of N-cadherin, Snail and Vimentin, while increased E-cadherin expression (p<0.05). After treatment with cisplatin, the expression of miRNA was further decreased after treatment of higher concentration of neo-gambogic acid.
Conclusion: Neo-gambogic acid can decrease miRNA106b and miRNA93 levels in Hela cells, increase cell sensitivity to cisplatin, increase E-cadherin expression, decrease N-cadherin, Snail and Vimentin expression, as well as inhibit Hela cell invasion and migration.Author(s): Feipeng Wang, Xiaoying Zhang