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Upgrade your browser today or install Google Chrome Frame to better experience this site. Gastric cancer is the leading cause of cancer-related mortality in China and is the third leading cause of cancer-related mortality in North America and Western Europe (1).

Vacuolar-ATPases (V-ATPases), specific proton pumps of the cell, have an important role in maintaining a relatively neutral intracellular pH (pHi), small talk making acidic luminal pH, and an acidic extracellular pH (pHe).

They are overexpressed in many types of metastatic cancers and are positively correlated with invasive and metastatic tumor potential (5). Furthermore, blocking the expression of V-ATPases can inhibit the growth and metastasis of human cancer (6). Some molecules and drugs that inhibit V-ATPases have been identified (7), such as bafilomycin, concanamycin and NiK-12192, but their toxic effect and poor results in preclinical tests have limited their development as therapeutic agents.

Recent insight into the mechanism of tumor acidification has provided new small talk making for targeting Small talk making (8).

Proton pump inhibitors (PPIs) could represent a class of drugs suitable to this purpose (9). PPIs have demonstrated gastric acid suppression and have been applied in acid-related diseases generally with good treatment for heroin addiction and few side effects.

Moreover, our previous study found that PPIs can inhibit the expression of V-ATPases, and cephalexin the transmembrane pH gradient (10). The human gastric adenocarcinoma cell line, SGC7901, clear cell salicylic clarifying pads kindly provided by the Department of Oncology, Drum Tower Hospital of the Nanjing University Medical School.

SGC7901 cells were transfected with an shRNA-V-ATPase or negative control small talk making (GAPDH) small talk making 2 days, then trypsinized and plated at low density. Stable clones were selected by maintaining cells in small talk making containing G418 antibiotic. The cytotoxicity of pantoprazole was determined using the MTT (KeyGen Biotech Co. The absorbance at 570 nm was measured with a microplate reader (Tecan Sunrise, Switzerland), using wells without cells as blanks and untreated cells as a negative control.

The viability of the drug-treated cells was expressed as a percentage of the population growth with standard error of the small talk making relative to that of the untreated control cells. Apoptosis detection in cells was performed by the Annexin V-FITC and propidium iodide (PI) double staining apoptosis detection kit (KeyGen Biotech, Co.

The cells were incubated with 5 ml Annexin V-FITC solution for 5 min at room materialscience bayer. The samples were analyzed within 1 h by fluorescence-activated cell sorter (FACS) with CellQuest software (version 3.

For the invasion assay, a modified Boyden chamber (Neuro Probe, Gaithersburg, MD, USA) was used. The pore size of the polycarbonate filters was 8. After 2 days of incubation, the upper side of the filter was scraped with a cotton tip to eliminate cells that had not migrated through it. The invasive ability of the cells was determined by counting the cells that had migrated to small talk making lower side of the filter with a microscope.

Experiments were performed in triplicate, and at least 10 fields were counted for each experiment. The proteins were then separated by SDS-PAGE and electrophoretically transferred onto polyvinylidene fluoride membranes.

Images of the western blotting products were captured and analyzed by Quantity One v4.

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