The Influence Of Vinorelbine On Growth And Apoptosis Of Esophageal Carcinoma Cells (Eca-109) In Vitro

Obj·ective:To study the influence of Vinorelbine on growth and apoptosis of esophagealcarcinoma cells (Eca-109) in vitro and to study the mechanism of the action of themedicine.Methods:1. The Eca-109cells were cultivated in vitro with Vinorelbine of differentconcentrations（2.5umol/L、5.0umol/L、10.0umol/L）. Inverted microscope was usedto observe the living cell morphology after48hours. After staining Eca-109cells byHE, observe the changes in cell morphology of each group.2. Analyze the influence of VINORELBINE of different concentraions（2.5umol/L、5.0umol/L,10.0umol/L）on the growth inhibition of Eca-109cells at differenttimes(12hours,24hours,48hours)with MTT and observe if there is any denpendenceon time or dose.3. Apply immunocytochemical method to mensurate the expression of Bcl-2and Baxafter the interference of VINORELBINE of different concentraions（2.5umol/L、5.0umol/L、10.0umol/L）on Eca-109cells.Results:1. Inverted microscopic analysis showed that Eca-109cells in the negative controlgroup grew adhering to cell walls, almost occupying the entire plate in a clear form.These transparent cells are irregular quadrangle or fusiform, showing the naturalextension. However, after the treatment of VINORELBINE of different concentraions,the growth of cells are obviously restrained compared to the negative control group. Intercellular space widened. The number of cells decreased compared to the negativecontrol group. These cells contracted. The rate of cell floating increased. The cellnuclei began to shrinkle. Apoptotic cells appeared, dead cells turning black, floatingand being adiaphanous.2. After stained by HE, under optical microscope, it could be observed that cells in thenegative control group were large, closely linked to each other, with various shapes ofcell nuclei. Nucleoplasm occupied a high percentage, pale. Nuclei enlarged, thephenomenon of nuclear fission appearing, with2-4nuclei. After the treatment ofVINORELBINE of different concentraions, the forms of cells changed, the propotionof nucleoplasm diminished, cells began to shrinkle, nuclear membrane split,chromatin concentrated, nuclear staining became thin and finally apoptotic cellsappeared.3. The testing result of MTT indicated that under the interference of VNB of the sameconcentration, the longer the action period was, the higher the inhibition ratio became.Time and the inhibition ratio were positively correlated. At the same time point, theinhibition ratio increased along with the rise of the concentration of VNB. Thus, theinhibition ratio and dose were also positively correlated. In all, the influence of VNBon the growth of Eca-109cells depended on dose and time.4. Immunocytochemical method was used to measure average optical density. Withthe concentration of VNB increasing, Bcl-2weakened and Bax graduallystrengthened.Conclusions:1. VNB can restrain the growth of Eca-109cells in vitro and its effect to some degreeis dependent on dose and time.2. VNB can control the apoptotic process by escalating the expression ofpro-apoptotic protein Bax and reducing the expression of anti-apoptotic proteinBcl-2.