The Effect Of QGS On The Migration And The Gap Junction Of Esophageal Cancer Cells

The first part: The effect of the QGS on the adhesion and migration of esophageal carcinoma cells.Objective:To investigate the enhanced effects of QGS on the cell-cell adhesion of the esophageal cancer cells, and reduced the adhesion between matrix and the esophageal cancer cells. Trying to confirmed the reduced effects of the QGS on migration of esophageal carcinoma cells via the effects of the adhesion.Methods:1 It is the first step of tumor metastasis that the adhesion between tumor cells is reduced and the tumor cells leave from the primary focus. This research selected TE1 which was the high differentiation of esophageal cancer cells and TE13 which was the low differentiation of esophageal cancer cells.Used MTT method and mechanical method to test the cell-cell adhesion of the esophageal cancers with QGS.2 This research selected TE1 and TE13. The effect of QGS on the adhesion between esophageal cancer cells and matrix was detected by electric cell-substrate impedance sensing.3 This research selected TE1 and TE13. Used wound scrape assay and electric cell-substrate impedance sensing to evaluated the cell motility and migration ability of esophageal carcinoma cells with QGS.4 This research selected TE1 which was the high differentiation of esophageal cancer cells and TE13 which was the low differentiation of esophageal cancer cells. The proliferation activity of esophageal carcinoma cells with QGS was detected by MTT assay.Results:1 MTT assay showed that the cell-cell adhesion of the cells with QGS(100?g/ml) enhanced significantly(P<0.05), and the result was the same as the mechancal counting result(P<0.05).2 Electric cell-substrate impedance sensing showed that adhesion of stroma and esophageal cancer cells with QGS(100?g/ml) was significantly lower than the control group(P<0.05).3 The wound scrape assay showed that the migration of esophageal carcinoma cell with QGS(100?g/ml) was significantly inhibited(P < 0.05).The result was confirmed by electric cell-substrate impedance sensing, which showed that the migration ability of the cells with QGS decreased significantly(P < 0.05).4 MTT assay demonstrated that no obvious effect of QGS on proliferation in TE1 and TE13(P>0.05).The second part: The effect of the QGS on the expression of Cx32 and the cytoskeleton rearrangement of the esophageal cancer cells.Objective: To investigate the enhanced effects of QGS on gap junction of esophageal carcinoma cells. To confirmed the inhibition of the Qigesan on migration of esophageal carcinoma cells, and to explore the possible molecular mechanism of Cx32 suppressing migration of esophageal cancer cells.Method:1 This research selected the high differentiation of esophageal cancer cells, TE1 and Eca109, the low differentiation of esophageal cancer cells,TE13. Used different concentrations of QGS(50?g/ml, 100?g/ml) to traet cells 24 h. Flow cytometry method and Western Blot were performed to evaluate the effect of expression of Cx32 in esophageal carcinoma cells with QGS.2 The expression of E-cadherin in esophageal cancer cells with QGS was tested by immunocytochemistry and flow cytometry.3 Esophageal cancer cells occur epithelial mesenchymal transition,pseudopodia come out from the cells and then gain migration ability. The morphological change of esophageal cancer cells with QGS were observed byimmunofluorescence assay.4 The change of microfilament cytoskeleton in esophageal cancer cells with QGS was observed by laser confocal microscopy.Results:1 Western blotting showed that the expression of Cx32 in esophageal carcinoma cells is closely related to the QGS concentration. The QGS(100?g/ml) could significantly enhance the expression of Cx32(P<0.05).Flow cytometry showed that in the cell membrane the expression of the Cx32 in esophageal cancer cells with QGS(100?g/ml) was significantly higher than the control group(P<0.05).2 Immunocytochemistry and flow cytometry results showed that the expression of E-cadherin in esophageal cancer cells with QGS was no significantly enhanced than the control group.3 QGS inhibits EMT of esophageal cancer cells was detected by immunofluorescence assay.4 The microfilament cytoskeleton in esophageal cancer cells with QGS arrayed more rules, while the microfilament cytoskeleton in the control group cells arrayed confusion. It is more obvious that the microfilament arrayed irregularly in the cell membrane.Conclusions:1 QGS can enhance the adhesion between esophageal cancer cells and inhibit the adhesion between esophageal cancer cells and the matrix.2 QGS has not effect on the proliferation of esophageal cancer cells in vitro environment, but can significantly inhibit the migration of esophageal carcinoma cells.3 QGS can inhibit the rearrangement of actin cytoskeleton in esophageal carcinoma cells.4 QGS can enhance the expression and function of Cx32, which may be the main mechanism of inhibition on cytoskeleton rearrangement and migration ability of esophageal cancer cells.