Experimental Research On The Effects Of Metformin Combinated With Celecoxib On The Proliferation And Apoptosis Of Human Lung Cancer Cell Line A549

Objective:Role of proliferation and apoptosis in human lung adenocarcinoma A549cells bymetformin and celecoxib, and to explore the underlying mechanisms.Methods:1.Metformin(1mmol/L、5mmol/L、10mmol/L), Celecoxib(25μmol/L) and the twocombined (1mmol/L+25μmol/L、5mmol/L+25μmol/L、10mmol/L+25μmol/L) weretreated cells, to observe the cell morphology under inverted microscope and to staincells by HE.2. To survey inhibition effect of Metformin and Celecoxib and the two combined inA549cells by MTT, and to observe concentration and time of Metformin whetherthere is a correlation.3. Metformin,Celecoxib and the two combined were treated lung cancer cell lineA549by immunocytochemistry method, then to observe the expression of Bcl-2andBax protein.Results:1. Using inverted microscope to observed A549cells. When the drug concentration ofmetformin group and combined group is increased, reducing the number of cells,decreasing the volume of cells, magnifying the gap of cells, shrinking nuclear of cells.Some cells appeared apoptosis. Using optical microscope to observed A549cells byHE staining. When the drug concentration of metformin group and combined group isincreased, decreasing the number of cells, changing morphological of cells, dissolving karyotheca of cells，lessening cytoplasm of cells. Some cell occurred apoptosis.2. MTT showed: The24h inhibition rate of A549cells were16.21±5.14,27.24±5.68,38.72±3.62,52.14±1.77,67.43±6.82,78.40±4.28,89.71±3.56in Metformin1mmol/L, celecoxib25μmol/L, metformin1mmol/L+celecoxib25μmol/L, metformin5mmol/L, metformin5mmol/L+celecoxib25μmol/L, metformin10mmol/L,metformin10mmol/L+celecoxib25μmol/L concentration; The48h inhibition ratewere17.17±2.84,31.43±4.23,47.13±4.34,64.81±1.25,76.53±4.24,83.57±6.76,93.15±2.64respectively; The72h inhibition rate were20.81±2.72,36.84±5.26,51.67±3.21,73.44±4.38,84.63±2.74,95.06±4.89,97.71±0.67. With the increase ofconcentration of metformin and metformin combibed celecoxib, the inhibitory ratewas raised, When the drug concentration constant, inhibition rate of cell increaseswith the extension of time. It has a concentration and time dependent.3. Immunocytochemistry showed: Metformin, celecoxib and metformin incombination with celecoxib were interventded A549cell. Expression of the Bcl-2protein was decreased, and expression of the Bax protein was increased.Conclusion:1．Metformin,Celecoxib and the two combined can obvious inhibit the appreciationof A549cells in vitro. It has a concentration and time dependent. When Metformincombined with Celecoxib, it has a more pronounced inhibition of proliferation inlung cancer cell line A5492. Metformin, celecoxib and metformin united celecoxib can cut through the Bcl-2protein and raised Bax protein. Making Bax/Bcl-2ratio increases, thus the cellapoptosis.