| Breast cancer,the number one tumor killer to threaten women in world today.The incidence and mortality of breast cancer is on the rise annually,the prevention and treatment of breast cancer are urgent nowadays.The effects and mechanism of anti-human MDA-MB-468 cells of breast cancer were studied with different categories Tieguanyin tea,tea polyphenols and theaflavins.Clean Tieguanyin tea,heavy Tieguanyin tea,stale Tieguanyin tea,dust of Tieguanyin tea and yellow leaves of Tieguanyin tea all according to the weight of tea and water volume ratio of 1:25 to extract.And fresh leaves of Tieguanyin tea according to the weight of tea and water volume ratio of 1:6.25 to tea polyphenols,EGCG and theaflavins.Then the sterilized tea was filtered as a drug to prevent the original concentration.To not add drugs as the normal control group.Such as EGCG,Tea polyphenols,theaflavins nature products of tea and 5-FU use prepare 1 mg/ml of drug intervention solution with PBS.The 5-FU drug was used as the positive control group and without adding the drug as the normal control group.Respectively breast cancer cell proliferation inhibition rate was detected by MTT assay,breast cancer cell apoptosis was detected by flow cytometry,and apoptosis-related proteins were detected by Western blotting,all of them to explore the effect and mechanism of inhibition of breast cancer.Finally,the content of main biochemical components of Tieguanyin was detected,and made with the results of different categories of Tieguanyin anti-breast cancer to take a correlation analysis.The results were showed as follows:1.The anti-cancer effects of different Tieguanyin teas on MDA-MB-468 cell proliferationThe effects of different categories of Tieguanyin tea on the cell proliferation inhibition rate of breast cancer MDA-MB-468 cells were showing dose-dependent relationship.At the same intervention concentration,the proliferation inhibition rate of breast cancer MDA-MB-468 cells was the highest in the stale Tieguanyin,and the inhibition rate of proliferation was the lowest in the fresh leaves of Tieguanyin tea.The concentration of IC50 in the proliferation inhibition rate of breast cancer MDA-MB-468 cells about different categories of Tieguanyin tea from big to small as follows:stale Tieguanyin tea(174.95 times)>heavy Tieguanyin tea(148.88 times)>of clean Tieguanyin tea(140.09 times)>yellow leaves of Tieguanyin tea(119.87 times)>dust of Tieguanyin tea(117.56 times)>fresh leaves of Tieguanyin tea(69.60 times).The six groups of Tieguanyin tea can clearly induce the apoptosis of MDA-MB-468 cells of breast cancer.Compared with the normal group,Percentage of apoptosis of MDA-MB-468 cells in the six groups of Tieguanyin tea was significantly increased(P<0.05)..And the highest rate of the apoptosis in the stale Tieguanyin tea,it's 28.53%.The fresh leaves of Tieguanyin tea was the lowest proportion of induced apoptosi,it's 18.53%.Sort by the proportion of apoptosis from high to low was:stale Tieguanyin tea>heavy Tieguanyin tea>fresh leaves of Tieguanyin tea>dust of Tieguanyin tea>yellow leaves of Tieguanyin tea>clean Tieguanyin tea.There were five apoptosis-related protein,which were the apoptotic proteins Bcl-2,Cleaved-caspase 8,Cleaved-caspase 9,Cleaved-caspase 3 and Caspase 3.Their expression levels were detected.Compared with the normal control group,the results were as follows:Clean Tieguanyin tea,heavy Tieguanyin tea,stale Tieguanyin tea,dust of Tieguanyin tea and fresh leaves of Tieguanyin tea can significantly decreased the expression of anti-apoptotic protein Bcl-2(P<0.05).Especially the inhibitory effect of the fresh leaves of Tieguanyin tea group was the most significant obvious,it's decreased 66.17%.The relative expression level of Cleaved-caspase 8 protein in MDA-MB-468 cells was significantly higher in clean Tieguanyin tea,heavy Tieguanyin tea,stale Tieguanyin tea,dust of Tieguanyin tea and yellow leaves of Tieguanyin tea(P<0.05).The relative expression level of Cleaved-caspase 8 in the clean Tieguanyin tea group was 80%higher than that in the normal group.Sort by the relative expression level from high to low was:clean Tieguanyin tea>dust of Tieguanyin tea>stale Tieguanyin tea>heavy Tieguanyin tea>yellow leaves of Tieguanyin tea>fresh leaves of Tieguanyin tea.The relative expression level of Cleaved-caspase 9 protein in MDA-MB-468 cells was significantly higher in fresh leaves of Tieguanyin tea,heavy Tieguanyin tea,dust of Tieguanyin tea and yellow leaves of Tieguanyin tea(P<0.05).The relative expression level of Caspase 3 precursor protein in MDA-MB-468 cells were significantly decreased by the six groups of categories Tieguanyin tea(P<0.05).But all the tea test groups to reduce the relative expression level of Caspase 3 precursor protein not more than 30%.The relative expression levels of Cleaved-caspase 3 were significantly higher in the six groups of Tieguanyin tea than in the normal group(P<0.05).All of he fresh leaves of Tieguanyin tea group,yellow leaves of Tieguanyin tea group and dust of Tieguanyin tea group were increased by more than 200%,and the lowest increase rate of the stale Tieguanyin tea group was 46%.Therefore,six different categories of Tieguanyin tea by inhibiting the expression of Bcl-2 protein,and promote the activation of Caspase 3,Caspase 8 and Caspase 9,thereby inducing MDA-MB-468 cells of breast cancer into the procedural death process,eventually induce cell apoptosis.This is an effective way to inhibit the proliferation of breast cancer MDA-MB-468 cells with these six different categories of Tieguanyin tea.2.The anti-cancer effects of tea polyphenols and theaflavins on MDA-MB-468 cell proliferationTea polyphenols and theaflavins on the cell proliferation inhibition rate of MDA-MB-468 cells have a significant inhibitory effectand,and showing dose-dependent relationship.At the same intervention concentration,the proliferation inhibition rate of breast cancer MDA-MB-468 cells was the highest in the 5-FU,and the inhibition rate of proliferation was the lowest in the theaflavins.When the intervention concentration reached 80ug/ml,the proliferation inhibition rateinhibitory rate of EGCG,tea polyphenols and anti-cancer agent 5-FU on MDA-MB-468 cells was more than 70%.The IC50 concentration of tea polyphenols and theaflavins on proliferation inhibition of breast cancer MDA-MB-468 cells was from low to high as follow:5-FU(19.51ug/ml)<EGCG(38.17ug/ml)<tea polyphenol(42.04ug/ml),theaflavin(75.08ug/ml).Tea polyphenols and theaflavins all could induce the apoptosis of breast cancer MDA-MB-468 cells(P<0.05).The apoptotic ratio of 5-FU group was 47.38%,while that of EGCG group was 31.08%.The apoptosis ratio induced by theaflavin group was the lowest,which was 20.66%.Sort by the induction ability from strong to weak was:5-FU group>EGCG group>tea polyphenols group>theaflavins group.There were five apoptosis-related protein,which were the apoptotic proteins Bcl-2,Cleaved-caspase 8,Cleaved-caspase 9,Cleaved-caspase 3 and Caspase 3.Their expression levels were detected.Compared with the normal control group,the results were as follows:5-FU,EGCG,tea polyphenol and theaflavin all could significantly decreased the relative expression level of Bcl-2 protein(P<0.05).The relative expression levels of anti-apoptotic protein Bcl-2 were sort from low to high was:theaflavins group<5-FU group<EGCG group<tea polyphenols group.The inhibitory effect of theaflavins group was the most obvious,down 57.72%.EGCG,5-FU,tea polyphenol and theaflavin all could significantly increased the relative expression level of Cleaved-caspase 8 protein(P<0.05).It's compared with that of normal group respectively increased by 3.83 times,3.56 times,2.74 times,2.42 times.5-FU,EGCG,tea polyphenol and theaflavin all could significantly increased the relative expression level of Cleaved-caspase 9 protein(P<0.05).Sort from high to low was:theaflavins group>tea polyphenols group>EGCG group>5-FU group.5-FU,EGCG,tea polyphenol and theaflavin all could significantly decreased the relative expression level of Caspase 3 precursor protein(P<0.05).The decreased amplitude of theaflavin group was the highest than that in the normal group,which was 40.28%.The relative expression levels of Cleaved-caspase 3 protein in 5-FU group,EGCG group,theaflavin group and tea polyphenol group were 1.98 times,1.91 times,2.20 times and 3.86 times higher than that of normal group.Therefore,the tea polyphenols and theaflavins have a strong ability to induce apoptosis of breast cancer MDA-MB-468 cells.On the effect of proliferation inhibition rate of breast cancer MDA-MB-468 cells and the ability to induce apoptosis,EGCG was the most effective,followed by tea polyphenols,and the effect of theaflavins was the worst.But the tea polyphenols and theaflavins in the study of the promotion and effect of breast cancer MDA-MB-468 apoptosis,showing a consistent trend,all by inhibiting the expression of anti-apoptotic protein Bcl-2,and promote the activation of pro-apoptotic proteins Caspase 8,Caspase 9 and Caspase 3,and then induce breast cancer cells apoptosis.3.The correlation analysis between the biochemical components from different Tieguanyin teas and their anti-cancer effectsThe results of biochemical composition analysis showed that the different categories of Tieguanyin tea had obvious effect on the apoptosis of breast cancer MDA-MB-468 cells.There was one or more projects had the extremely relevant relationship between the biochemical composition and IC50 of breast cancer inhibition rate,apoptosis rate,the expression of Cleaved-caspase 9,Cleaved-caspase 3 and Caspase 3(|r|>0.8).Especially the total amount of catechins,plays an important role in the expression and activation of Caspase 3 and the expression of Cleaved-caspase 3.The results together illustrate the various components of tea water extracts,can co-inhibit the proliferation of breast cancer cells.Combined the effect of tea extracts to anti-breast cancer MDA-MB-468 cells can be found,EGCG,theaflavin and tea polyphenols also have a good effect on inhibit the proliferation of cancer cells in unilateral.And all in the IC50 concentration of the intervention,the three kinds of tea extracts to show the ability to induce apoptosis is not inferior to the different categories of Tieguanyin tea.Which EGCG had a higher proportion of induced apoptosis ratio,is more than the different categories of Tieguanyin tea.It shows the strength and advantage of effect in extracts.But it needs to be at a higher concentration to play a good anti-cancer effect. |
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