| Objective: To explore the effects of fig leaf extract on the biological activities of colon cancer cells such as proliferation,migration,apoptosis and cell cycle and the possible related mechanisms of action.Methods:Clinical theoretical study: deep reading ancient books and modern literature,the colon cancer of traditional Chinese and western medicine clinical theory and related description,etiology,clinical manifestations,diagnosis,syndrome differentiation is summarized,and the pharmacological effect of fig leaf extract conclude,provide theoretical basis for further research.Experimental Research:Colon cancer cell lines HT29 and SW480 were cultured in vitro with different concentration gradients(10 μM,30 μM,50μM)of bergamot lactone,and the effects of different concentrations(10 μM,30 μM,50 μM)of bergamot lactone on the proliferation and invasion of colon cancer HT29 and SW480 cells were examined by CCK-8 assay,Ed U assay,cell clone formation assay,scratch assay,Transwell assay,flow cytometry assay,cell cycle assay,The effects of different concentrations(10 μM,30 μM and 50 μM)of bergamot lactone on the proliferation,invasion,migration,apoptosis and cycle changes of colon cancer HT29 and SW480 cells were examined by CCK-8 assay,Ed U assay,cell clone formation assay,scratch assay,Transwell assay,flow cytometry assay,cell cycle assay and Western-blot assay.The obtained results were statistically processed and analyzed to describe the experimental data its measurement results as mean ± standard deviation.For data conforming to normal distribution,independent samples t-test was used to compare the significant differences between the two groups of data.Results:Clinical theoretical study:(1)The traditional Chinese medicine name of colon cancer is "intestinal accumulation","dysentery","intestinal wind","dirty poison","lock anal hemorrhoids" and so on.Its etiology pathogenesis is due to is evil,gas,poison,stagnation in the colon,over time,early is given priority to with evil reality,late more for vital qi deficiency or false inclusion,heat,poison,stasis,phlegm,this virtual mark,treatment should be both specimens,dispel evil and upright weight,cure,dredge qi,promoting blood circulation,heat clearing clearing dampness.(2)Fig leaf is an ancient Chinese medicine,with intestines and stomach,heat and detoxification,swelling and pain relief,can treat enteritis,damp and heat diarrhea,dysentery,belt,hemorrhoids,carbuncle pain,furuncle scabies and other diseases.Its active ingredients also have a role in anti-inflammatory,anti-viral,anti-tumor,anti-oxidation,improving gastrointestinal function,improving immunity,lowering blood lipid and blood sugar.As one of the main extracts of fig leaves,many scholars have proved that it has good efficacy in anti-tumor,and its mechanism of action may have an inhibitory effect in the proliferation,invasion,migration and apoptosis of malignant tumors.Experimental Research:(1)Compared with the control group,the proliferation of colon cancer cells HT29 and SW480 was inhibited by bergamot lactone dosing at different concentrations(10 μM,30 μM and 50 μM)(p<0.05),and the inhibition rate increased with the increase of drug concentration;(2)The results of Ed U assay also confirmed that bergamot lactone could inhibit the proliferation of HT29 and SW480 cells in a concentration-dependent manner(p<0.05).(2)Ed U assay also confirmed that bergamot lactone could concentration-dependently inhibit the proliferation of HT29 and SW480 cells(p<0.05);(3)Clonogenesis assay showed that bergamot lactone inhibited the clonogenesis of colon cancer cells(p<0.05);(4)Scratch assay showed that bergamot lactone could inhibit the migration of colon cancer cells(p<0.05).(5)Traswell showed that bergamot lactone inhibited the invasion of colon cancer cells(p<0.05);(6)Flow cytometry assay showed that different concentrations of bergamot lactone could induce apoptosis in two types of colon cancer cells(p<0.05);(7)bergamot lactone could increase the percentage of G1 stage cells(p<0.05).(8)Bergamot lactone could increase the expression of pro-apoptotic proteins and decrease the expression of inhibitory apoptotic proteins in colon cancer cells(p<0.01).Concusion: According to the symptoms colorectal cancer probably belongs to the category of "intestinal accumulation","dysentery","intestinal wind","dirty poison" and "locked hemorrhoids" in traditional Chinese medicine."Dysentery","intestinal wind","dirty poison","locked hemorrhoids" and other diseases.Mostly due to weakness of the spleen and stomach,poor diet,emotional disorders,blood stagnation and other factors leading to the imbalance of Qi and blood in the body,damage to the internal organs,dampness and heat,blood stagnation in the colon accumulation.The treatment should take into account both the symptoms and the root cause,and the treatment should focus on eliminating the evil and helping the righteous,as well as unblocking the qi,activating blood circulation and removing blood stasis,clearing heat and detoxifying dampness.As one of the main extracts of fig leaves,this study showed that bergamot lactone could effectively reduce the viability of human colon cancer HT29 and SW480 cells,increase the percentage of G1 phase cells,and cause cell cycle arrest.It also promoted apoptosis of colon cancer cells by regulating the expression of apoptosis-related proteins PARP,BAX,and Bcl-2. |
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