CYTOTOXIC EFFECT OF TAXOL AND VITAMIN E SUCCINATE COMBINATION IN COMPARISON WITH TAXOL ALONE ON MCF-7 BREAST CANCER CELL CULTURE BY MTT ASSAY

3. Results:-
Absorbance of all the 3 plate well solutions for all the 3 experiment ( 3 drug regimen to be tested ) were measured and recorded in the tables below.

Cytotoxicity of Taxol  and Vitamin E succinate:-  We concluded dose and time –response studies on the on the proliferation of MCF-7 breast cancer cells in vitro to determine the effects of Taxol and VES on breast cancer growth. After 72 hr of incubation, both Taxol and VES inhibited cell growth in a dose-dependenmanner in MCF-7 breast cancer cell lines which can be cleared from figure 5 (A, B, C)

Absorbances recorded for taxol are shown in table below

(Table 1):-Measured absorbance(cell viability) and % cell viability for Paclitaxel at 570nm in spectrophotometer  of a MTT assay drug solution for all three 24 well plate)

( Table 2) :-  Measured absorbance(cell viability) and % cell viability for Vitamin E Succinate  at 570nm in spectrophotometer  of a MTT assay drug solution for all three 24 well plate )

Measured absorbance of the MTT assay final purple colour solution indicates cell viability after treatment with drug. So, we can say that it is the indirect measure for cytotoxicity of drug. Percentage cell viability will give us more clear idea to compare cytotoxicity of different drug regimen because IC50 of drug is much easier to calculate from graph of % cell viability Vs concentration. IC 50 is the half maximal inhibitory concentration and is a measure of effectiveness of a compound in inhibiting biological function. This measure indicates how much of a particular drug is needed to inhibits a given biological process by half ( Wikipedia IC50, 2010).The below figures are the graphs for % cell viability vs Concentration.

(Figure 5 (A) :-  Cell viability measured by MTT assay for Paclitaxel. It shows cytotoxicity  of paclitaxel on MCF-7 cell lines which are treated with different concentration described in table 1 of paclitaxel for 72 hrs. Graph is % cell viability against concentration of taxol. )

(Figure 5 (B) :- Cell viability measured by MTT assay for VES. It shows cytotoxicity of Vitamin E succinate on MCF-7 cell lines which are treated with different concentration described in table 2 of VES for 72 hrs. Graph is % cell viability against concentration of VES. )

From the above graph, it is clear that IC50 for paclitaxel is 8.9 nM and IC 50 for Vitamin E Succinate is 25µM. This IC50 of VES was used for the taxol-VES combination regimen experiment.

Eventually, IC 50 of all the 3 regimen ( Taxol, VES, Taxol-VES combination) can be compared to find out most effective regimen for MCF 7 breast cancer cell i.e lower the IC 50 of the drug more effective the drug in comparison to other

Cytotoxicity of Taxol- VES combination:- The measured absorbances of the MTT assay for this Taxol-VES combination are recorded in table below

( Table 3:-   Measured absorbance(cell viability) and % cell viability for Paclitaxel + 25µM Vitamin E Succinate (IC50 of VES)) at 570nm in spectrophotometer  of a MTT assay drug solution for all three 24 well plate.

(Figure 5 (C) :-  Cell viability measured by MTT assay for Taxol-VES combination. It shows cytotoxicity of paclitaxel-Vitamin E succinate on MCF-7 cell lines which are pre- treated with IC50 concentration of VES (25µM) and then treated with different concentration described in table 1 of paclitaxel for 72 hrs. Graph is % cell viability against concentration of taxol-VES combination. )

From the figure 5(C) IC50 of taxol- Vitamin E succinate combination is 2 nM.

Now, it is time to move on our hypothesis of our study i.e to compare cytotoxicity of taxol and taxol-VES combination and it can be cleared by comparing % cell viability vs concentration graph for both regimen ( taxol and taxol-VES combination ) which is shown below. Figure 6  is the most important figure to conclude our study aim and it is discussed in discussion section of this work.

( Figure  6:  Comparative cytotoxic effects of taxol-VES combination therapy and taxol therapy alone on MCF-7 breast cancer cell lines treated for 72 hrs. The graph is %cell viability against concentration for appropriate drug therapy )

Independent Sample T-test:-  The raw data result format is submitted in appendix. Levene’s test for equality of variances is not significant because P-Value is 0.426 >>>0.05. So, t-test can be applied to given data set. Q-Q plot shows normal distribution of data. Box plot has no outliers so data is not skewed i.e it is normally distributed which is required for any statistical analysis.

Hypothesis for the test is there is no significant difference between these two therapy ( taxol-VES combination and Taxol alone). i.e mean value of cell viability (absorbance)for both therapies are same μ1=μ2. P value for T-test at 95 percentage of confidence interval is P=0.02<<<0.05. So, our hypothesis for the test is rejected which indicates there is significant difference between these two drug therapy. More specifically, it is analysed in discussion.

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