RNA interference targeting ATP6L of breast cancer cells in vitro inhibition of invasion.doc

 PAGE \* MERGEFORMAT 21 RNA interference targeting ATP6L of breast cancer cells in vitro inhibition of invasion Study: Haiyan Yu, Jie Jin, Downing, Yun Deng, Shu Wai Kwan, Shen Qiu Jin, QIN Wen New [Abstract] Objective: RNA interference targeting ATP6L the proton pump inhibiting the function of the detection of breast cancer cells MCF 7/ADR invasion. Methods: The effective ATP6L interference and interference control fragments were transfected into MCF 7/ADR cells by BCECF fluorescence spectrophotometer with cells incubated with supernatant excited at different excitation the fluorescence emission; by real-time fluorescence quantitative PCR detection of cells in the matrix metalloproteinase 2 (MMP 2) expression; method with gelatin zymography Detection of cells in the MMP 2 supernatant activity; by cell invasion assay (Transwell) to detect cell invasion in vitro. Results: MCF 7/ADR cells, the expression of genes down ATP6L cells can secrete H + diminished capacity; cells MMP 2 expression level did not change; cell secretion of MMP 2 supernatant activity decreased; cells decreased invasive ability in vitro. Conclusion: The RNA interference targeting ATP6L inhibit the function of the proton pump, thereby inhibiting breast cancer cells MCF 7/ADR in vitro invasion. [Keywords:] ATP6L; RNA interference; invasion; tumor [Abstract] Objective: To explore whether the invasion of MCF 7/ADR cells could be affected by in vitro inhibition of V ATPase activity. Methods: The expression of ATP6L was knocked down by specific siRNA in breast cancer cell line, MCF 7/ADR . Then the capability of proton extrusion was analyzed by detecting the extracellular pH of the supernatant using pH sensitive fluorescent dye BCECF on a luminescence spectrometer. The expression of MMP 2 and activity of gelatinase were determinated by real time fluorogentic quantitative PCR and zymography. The ability of cell invasion in vitro was tested by transwell assay. Results: Proton extrusion was