ABSTRACT STUDY OF MOLECULAR BIOLOGY THE EXPRESSION OF P53, CYCLIN D1 AND BAX IN BREAST CANCER CELL LINE T47D AFTER TREATMENT OF MUCOXIN

 

Muhartono

Mucoxin is one of acetogenin which is isolated from leaves of Rollinia mucosa, can be used as an anti‒tumor agent, believed to work by inhibiting proliferation and increase apoptosis, and served to increase the expression of p53 and Bax and decreased expression of cyclin D1. The aim of this research is to identify mucoxin effect on the expression of genes p53, Bax and cyclin D1 in breast cancer cell line T47D.

This study was analytic with cross sectional approach,  and purely experimental which is done on 72 groups. Samples were divided into 2 groups, one group were not given mucoxin, and the test group were given mucoxin 1×10‒3µg/ml; 0.5×10‒3 µg/ml; 1×10‒3 µg/ml; 5×10‒3µg/ml; 10×10‒3µg/ml, performed 3 repetitions examination. Expression of p53, Bax and cyclin D1 gene were examined using RT‒PCR, protein expression of p53, Bax and cyclin D1 were examined using immunohistochemical techniques and the value of apoptosis and proliferation checked using flow cytometry. Samples examined at  0, 24, 48and 72hour after exposure mucoxin, compared with controls. Data were tested statistically using Anova, Kruskal Wallis, Mann Whitney, simple linear regression and multiple linear regression.

Result show there is an impairment of proliferation and increase in apoptosis values ​​with increasing dose and exposure time of mucoxin. Kruskal Wallis test results in proliferation examination at 0, 24, 48 and 72 hours, obtained p value=0.007, 0.009, 0.006 and 0.006. Kruskal Wallis test results in apoptosis examination at 0, 24, 48 and 72 hours, obtained p value=0.008, 0.012, 0.005 and 0.005. There are differences in the expression of p53, cyclin D1 and Bax in cell line T47D were given mucoxin compared with those not given mucoxin. Results of Kruskal Wallis and Anova test in p53 expression at 0, 24, 48 and 72 hours, obtained p value=0.000, 0.008, 0.000 and 0.005. Results of Kruskal Wallis and Anova test in cyclin D1 expression at 0, 24, 48 and 72 hours, obtained  p value = 0.0001, 0.0001, 0.0001 and 0.006. Results of Kruskal Wallis and Anova test  in Bax expression at 0, 24, 48 and 72 hours, obtained p value=0.007, 0.011, 0.0001 and 0.012. There is a relationship between p53 expression with the value of proliferation and apoptosis in cell line T47D were given mucoxin. Result of simple regression analysis of p53 expression with apoptosis value, obtained p value=0.000 with the positive correlation. Result of simple regression analysis of p53 expression with proliferation value, obtained p value = 0.000 with a negative correlation. There is effect of p53 expression in Bax and cyclin D1expression. Results of multiple linear regression analysis of p53 expression with Bax and cyclin D1expression, obtained p value=0.004.

Conclusions, There are differences in the expression of p53, cyclin D1 and Bax in cell line T47D given mucoxin with control. There is a relationship between the expression of p53, cyclin D1 and Bax with proliferation and apoptosis cell line T47D. There is effect of p53 expression on the expression of cyclin D1 and Bax.

 

Key word: Apoptosis, Bax, Cyclin D1, Mucoxin, P53, Proliferation, T47D.

 

Abstrak Muhartono

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