Cytotoxicity, Regulation of Apoptotic and Anti-apoptotic Gene Expression by IL-27 in MCF-7 and MDA-MB-231 Breast Cancer Cell Lines

Abstract

Breast cancer is one of the commonest cancers among women. Conventional therapies cause adverse side effects in patients. Cytokine immunotherapy such as has been sought as an alternative cancer treatment in recent years. has been shown to improve anticancer immunity and anti-angiogenesis in cancers, however, its effect on apoptotic and anti-apoptotic gene expression especially in breast cancers is yet to be explored. Cytotoxicity of and cancerous breast cell lines was first determined for 24-72 h in this study. The results indicated that IL-27 treatment did not retard 184b5 cell growth, however, did inhibit and respectively. Apoptotic (TRAIL, FADD, FAS, and anti-apoptotic (BCL-2, AKT, and COX-2) genes were then amplified from untreated (control) and treated breast cancer cells and studied. TRAIL, caspase-3, caspase-8 gene expression was significantly (p < 0.05) upregulated in treated MCF-7 (442 ng/ml) and MDA-MB-231 (457 ng/ml) cells. Expression of FADD and FAS genes was not detected in both control and treated MCF-7 and MDA-MB-231 cells. COX-2 gene was also not expressed by MCF-7 cells, but reduced significantly (p < 0.05) in treated MDA-MB-231 cells. In MDA-MB-231 cells, IL-27 treatment seemed to slightly enhance the expression of AKT and BCL-2 genes which, on the other hand, was downregulated in treated MCF-7 cells. Conclusively, IL-27 is able to inhibit breast cancer cell growth and regulate apoptotic and anti-apoptotic gene expression in breast cancer cells. Keywords: IL-27; cytokine immunotherapy; triple negative breast cancer; invasive ductal breast cancerABStRAK Kanser payu dara adalah antara kanser yang paling biasa dihidapi oleh wanita. Rawatan konvensional menyebabkan kesan sampingan teruk dalam pesakit. Imunoterapi sitokin seperti interleukin-27 (IL-27) telah digunakan sebagai rawatan alternatif sejak kebelakangan ini. IL-27 telah terbukti dapat menambah baik imuniti antikanser dan aktiviti anti-angiogenesis dalam kanser, namun kesannya pada pengekspresan gen apoptotik and anti-apoptotik terutamanya pada kanser payu dara masih belum diterokai. Dalam kajian ini, kesan sitotoksisiti IL-27 pada sel selanjar bukan kanser (184b5) dan kanser (MCF-7 dan MDA-MB-231) ditentukan pada 24-72 jam dalam kajian ini. Dapatan kajian menunjukkan rawatan IL-27 tidak menjejaskan pertumbuhan sel 184b5, namun menghalang pertumbuhan sel and FADD, FAS, caspase-3 dan caspase-8) and anti-apoptotik (BCL-2, AKT, dan COX-2) kemudiannya diamplifikasikan daripada sel kanser payu dara kawalan dan terawat serta dikaji. Pengekspresan gen TRAIL, caspase-3 dan caspase-8 telah meningkat secara signifikan (p < 0.05) dalam sel yang dirawat. Pengekspresan gen FADD dan FAS gagal dikesan dalam sel MCF-7 dan MDA-MB-231, mahupun sel kawalan atau terawat. Gen COX-2 tidak diekspreskan oleh sel MCF-7 tetapi pengekspresannya telah menurun secara signifikan (p < 0.05) dalam sel MDA-MB-231 yang dirawat. Dalam sel MDA-MB-231, rawatan IL-27 meningkatkan pengekspresan gen AKT dan BCL-2, namun pengekspresan kedua-dua gen tersebut telah menurun...