Effects Of Different Types Of Opioids On Proliferation And Migration Of Lung Cancer Cell Line A549

Objective: By studying effects of morphine,sufentanil and butorphanol on proliferation,invasiveness and migration of lung carcinoma cell line A549 respectively,we want to provides a theoretical basis for the use of opioids in the treatment of lung cancer.Methods:(1)Opioid receptor expression was measured in human lung cancer cell line NCI H358,NCI-H1975 and A549 with Western Blot.(2)Proliferation of A549 cell line was decreased 24 hrs after medication with different concentrations of morphine(0.1μmol/L,1μmol/L and 10μmol/L),sufentanil(0.1nmol/L,1nmol/L and 10 nmol/L)and butorphanol(0.1μmol /L,1μ moe /L and 10 μmol/L)respectively.(3)Proliferation of A549 cell line was tested by using clone formation experiment 7 days after medication with different concentration of morphine(0.1μmol/L,1μmol/L and 10μmol/L),sufentanil(0.1nmol/L,1nmol/L and 10 nmol/L)and butorphanol(0.1μmol/L,1μ mol/L and 10μmol/L)respectively.(4)culture A549 cells in transwell chamber covered with Matrixgel.Invasiveness of A549 cells was detected via transwell experiment,24 h after medication with different concentration of morphine(0.1μmol/L,1μmol/L,10μmol/L),sufentanil(0.1nmol/L,1nmole/L,10 nmol/L)and butorphanol(0.1μmol/L,1μmole/L,10μmol/L)to A549 cells respectively.(5)Migration rate of A549 cell line was measured by wound healing assay,24 h after medication with different concentration of morphine(0.1μmol /L,1μmol/ L,10μmol/L),sufentanil(0.1nmol /L,1nmol/L,0nmol/L)and butorphanol(0.1μmol/L,1μmol/L,10μmol/L)to A549 cells respectively.Result: the A549 cells also expressed the μ receptor(MOR)and the κ receptor(KOR).(2)the MTT test results showed that morphine decreased the proliferation of A549 cell line when the concentration was 10μmol/L,but sufentanil and butorphanol did not affect the proliferation of A549.(3)Cell cloning experiment result showed that morphine decreased the number of A549 cells when the concentration was 10 μmol/L,but sufentanil and butorphanol did not affect the number of sets of A549.(4)The results of transwell experiment showed that,when the concentration of morphine was 0.1μmol/L,and 1μ mol/L,the invasion rate of A549 cell line was increased,but the invasion rate of A549 cell line decreased when the concentration of morphine was 10μmmol/L.As the concentration of sufentanil increased,the invasion rate of A549 cell line increased.As the concentration of butorphanol increased,the invasion rate of A549 cell line decreased.After adding the corresponding antagonist,the effect of the opioid on the invasion ability of A549 was eliminated.(5)The results of wound healing assay showed that when the concentration of morphine was 0.1μmol/L,migration rate of A549 cell line was decreased,but when the concentration of morphine was increased to 1μmol/L and10μmol/L migration rate of A549 cell line was increased.As the concentration of sufentanil increased,the migration rate of A549 cell line was increased.As the concentration of butopenol increased,migration rate of A549 cell line was decreased.After adding the corresponding antagonist,the effect of the opioid on the migration rate of A549 cell line was eliminated.Conclusion:⑴When the concentration of morphine was 10μmol/L,the proliferation and migration of A549 cell was inhibited.When the concentration of morphine was 0.1μmol/L,the A549 cell migration was promoted.⑵ Sufentanil had no effect on the proliferation of A549 cell but promoted migration of A549 cell.⑶Butorenol had no effect on the proliferation of A549 cell and inhibited migration of A549 cell.