Evaluating the effects of Cucurbitacin D on Apoptotic and Autophagic genes expression in breast cancer cell line MDA-MB 468

Introduction: Breast cancer is the most common malignancy among females affecting approximately one out of ten women. Cucurbitacins are highly oxidized tetracyclic triterpenoids has been reported to have anticancer effects and can induce variety of programmed cell death signaling pathway. We examined the effect of Cucurbitacin D on apoptotic and autophagic genes expression in breast cancer cell line MDA-MB 468.Method: Gene expressions were analyzed by Real Time PCR. Total RNA was isolated from a series of MDA-MB 468 cells by RNeasy Mini Kit of QIAGEN. First strand cDNA was synthesized from extracted RNA using an Oligo dT as the primer. After cDNA was synthesized, the primers of target genes were employed and QuantiFast™ SYBR Green PCR master mix was used to amplify the genes. Apoptotic genes such as Bax, Bcl2, Casp3, p53 and vital autophagic genes such as Atg5, Beclin1and LC3 were studied by Real Time PCR.Result and Conclusion: The result demonstrated that cucurbitacin D can induce autophagy pathway in MDA-MB 468 through Atg5 dependent mechanism by overexpression of LC3 and Atg5genes. Also cucurbitacin D can inhibit apoptosis pathway by upregulation of anti-apoptic Bcl2 gene and downregulation of proaptotic Bax and Casp3 genes.