Synergistic Effect Of Sesamin And Gemcitabine On Human Lung Cancer Cell Lines A-549Influence

Objective:To observe the effect of sesamin and sesamin in conjunction with gemcitabine on human lung cancer cell lines A-549.Methods:The human lung cancer cell lines A-549were cultured in vitro by MTT colorimetric determination to determine the anti-proliferation effect of sesamin, gemcitabine, and sesamin in combination with gemcitabine on human lung cancer cell lines A-549.Morphological changes of cell apoptosis was observed after sesamin, gemcitabine, and sesamin in combination with gemcitabine effected on human lung cancer cell lines A-549by inverted microscope and the fluorescence staining of Hoechst33258.Concentration change of intracellular Ca2+was quantitatively analysised and detected by flow cytometry. The expression of caspase-3,8,9of the human lung cancer cell lines A-549was observed by opital microscope after the use of Immunocytochemical stain.Results:①The MTT colorimetric results showed that low concentration of sesamin had a certain depressant effect on human lung cancer cell lines A-549,and gemcitabine of various concentrations had the inhibition on human lung cancer cell lines A-549and with the increase of drug concentration the trend of the inhibition ratio heightened, and the inhibition ratio of sesamin uniting gemcitabine was more significantly enhanced than the separate application of gemcitabine on the cell lines A-549,P <0.01,so difference was statistically significant;②Inverted microscope and fluorescence staining of Hoechst33258were used to carry out cell morphological observation,it could be found the number of adherent cells was lesser in the sesamin group, gemcitabine group and combination group than in the control group,and the cells could not adhere strongly,or off the wall to float.and some cells appeared smaller in size, to offer small round globular,and cell-cell junction disappeared, then the break-off phenomenon could be seen among the surrounding cells.At the same time, the cell debris, loss of the original polygonal or spindle-shaped morphous of the tumor cell could be found, so tumor cell growth was inhibited,and compared with gemcitabine group, cell morphology of the combination group changed more significantly. The changs included karyopyknosis, nuclear chromatin high degree of cohesion, and marginalization.Compared with single-use gemcitabine, performance of the combination group was more obvious;③After48hours effect of drug, compared with early apoptosis ratio of the control group, sesamin group, gemcitabine group and combination group increased more obviously by the flow cytometry detecting early apoptosis ratio of cells, and compared with early apoptosis ratio of gemcitabine group,combination group increased more significantly[control group (3.75±0.10%),sesamin group (12.40±0.52%), gemcitabine group (13.32±0.71%); the combined group(26.59±0.76%).sesamin group, gemcitabine group respectively compared with the control group, p<0.05; the combination group conmpared with control group, P<0.01sesamin group compared with gemcitabine group, p=0.71; combined group compared with gemcitabine group, p<0.01)];④After48hours effect of drug, compared with concentration level of intracellular Ca2+of the control group, sesamin group, gemcitabine group and combination group increased more obviously by the flow cytometry detecting concentration change of intracellular Ca2+,and compared with of early apoptosis ratio of gemcitabine group, combination group increased more significantly [control group (2.16±0.11), sesamin group (12.11±0.67), gemcitabine group (15.46±0.97), the combination group (54.72±3.56); sesamin group, gemcitabine group respectively compared with the control group, p<0.05; the combined group conmpared with control group, P<0.01; combination group compared with gemcitabine group,p<0.01];⑤After48hours effect of drug, compared with expression of Caspase-3,8,9in the control group,sesamin group,gemcitabine group and combination group increased more obviously by the fluorescence staining of Hoechst33258, and compared with the expression of Caspase-3,8,9of gemcitabine group, combination group heightened more significantly [the expression of Caspase-3in the control group (8.31±0.49%), sesamin group,21.81±0.67%gemcitabine group (46.97±3.26%), the combination group(61.48±1.04%);sesamin group,gemcitabine group singlely compared with the control group, p<0.01combination group compared with gemcitabine group, p<0.01];[expression of caspase-8in the control group,(6.56±0.76%), sesamin group (20.73±0.55%), gemcitabine group (48.16±1.27%), the combination group (57.72±1.19%);sesamin group, gemcitabine group respectively compared with the control group,p<0.01combination group compared with gemcitabine group, p<0.05];[expression of Caspase-9in the control group,(7.21±1.25%), sesamin group (24.73±1.53%), gemcitabine group (49.12±1.17%), the combination group (59.45±0.93%); sesamin group,gemcitabine group respectively compared with the control group,p<0.01;the combination group compared with gemcitabine group, p<0.05)].Conclusion:(1) Low concentration of sesamin on human lung cancer cell lines A-549has a certain inhibitory action;(2) Sesamin uniting gemcitabine effect on lung cancer cells shows a significant inhibition,and a good synergistic role of anti-malignant tumor cells;(3)Sesamin can induce apoptosis by elevating intracellular calcium concentration of lung cancer cell, also strengthen the effect of gemcitabine that can enhance the calcium concentration in the cytoplasm of lung cancer cell to induce apoptosis;(4)Sesamin not only participates activation of Caspase pathway,but also enhances the ability that gemcitabine activates Caspase family to induce tumor cell apoptosis.