Identifying the cytotoxic role of Cassia fistula extract on LS174T cell line via modulation apoptotic and inflammatory markers

Abstract

Back ground

Many medicinal plants, including Cassia fistula, contain flavonoids, which have anti-inflammatory properties because the plant species are rich in antioxidant compounds. Numerous functions of flavonoids have been demonstrated, including the inhibition of numerous enzymes that are triggered during the inflammatory process.

Aim of study

Identify the cytotoxic impacts of C. Fistula in LS174T cell line colon cancer by affecting the apoptotic and inflammatory markers.

Material and methods

LS174T cell line was seeded in 96-well plate for exposure six serial dilution of in order to produce a stock solution of C. Fistula, was prepared and the seeded LS174T cell line was exposed to different concertation of C. Fistula (250,125,.62.5,31.2,15.6, 7.8μg/ml). The study design consists of control untreated group that involved LS174T cell line that subjected to RPMI1640 only and treated group that subjected to mentions concentrations of C.fistula. At then incubated for 48 hours. MTT assay was perform for estimation the cytotoxic effect of the Fistula on LS174T cell.  For evaluation apoptotic markers (Caspase-3and MCL-1) and inflammatory markers (IL-1B and IL-10), the cells were collected by adding trypsin to detach the cells, for getting the supernatant, and keeping the cells in a 1.5 ml Eppendorf tube for By employing the Sandwich-ELISA methods.

Results

 When compare to control group, All the concentrations of C. fistula archived significant reduction in the viability LS174T colon cancer cell line in dose dependent manner P<0.01. The highest concentration of C. Fistual (250,125,.62.58, 15.6 μg/ml) causes highly significant(P<0.001) elevation in Caspase-3 level among the control while lower concentration shows non-significant (P>0.5) effect on level of caspase-3 in compare to control. MCL-1 level in colon cancer cells significantly declined at higher three concentrations of C. fistula treated cells in comparison to the control group. On the contrary, there was no significant difference at the lower three concentration of Cassia Fistula extract. there was a significant decline in IL-1β level for all concentrations except with the lowest one when compared with the control group. as compared to the control group, the IL-10 level significantly elevated at all concentrations of C.fistula  except the lowest one.

Conclusion

Cassia Fistula extract can inhibit colon cancer cell proliferation, promote cell apoptosis, and control tissue inflammation, indicating anticancer activity.