Introduction: Breast cancer is one of the leading cause of cancer deaths in women. Metastasis in BC is caused by immuno- surveillance deficiency, such NK cell maturation, low NK activity and decreasing cytotoxicity. This study was performed to improve activating receptors and cytotoxicity of NK cells using interleukins (ILs). Methods: Human recombinant IL-2, -15, and -18 were used to induce NK cells. We measured the activating and inhibiting receptors, proliferation activity of NK cells, and the cytotoxicity of NK cells on BC cells (MCF7). The effects of ILs were tested on the NK cell receptors CD314, CD158a and CD107a with flowcytometry, proliferation at various incubation times with 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy methoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and concen- trations of TNF-α and IFN-γ by NK cells with ELISA. Results: ILs increased NK cell receptor levels (CD314, CD158a, and CD107a) at 24 hours of incubation. ILs increased NK cell viability, which increased with longer incubation. Moreover, ILs-induced NK cells inhibited proliferation in MCF7 cells, as well as increased TNF-α, IFN-γ, PRF1 and GzmB secretion. Conclusion: IL-2, IL-15, and IL-18 improved activating receptors and proliferation of NK cells. IL-induced NK cells in- creased TNF-α, IFN-γ, PRF1 and GzmB secretion and cytotoxic activity on BC cells. High NK cell numbers increased BC cell growth inhibition. Keywords: Activator; breast cancer; interleukins; natural killer; receptor.

Breast cancer (BC) is the most prevalent type of cancer among women and one of the major causes of cancer mortality in women. Metastasis in breast cancer (BC) occurs due to immunosurveillance deficiency, including impairment of natural killer (NK) cell maturation. Conditioned medium (CM) from human Wharton's jelly mesenchymal stem cells (hWJMSC-CM) is known to possess anticancer activity. The CM of co-culture of human recombinant IL-2 treated NK cells and hWJMSCs is expected to boost anticancer activity toward BC cells which can be analyzed from the effect of CM towards secretion of effector molecules and expression of BC cell apoptosis-related genes, and cytotoxic granules in human recombinant IL-2 treated NK (IL-2 NK) and hWJMSCs (IL-2 hWJMSCs). TNF-α, IFN-γ, perforin, granzyme were measured by ELISA, while the inhibition of cell proliferation was measured by MTS assay and BC cell apoptosis by flow cytometry and apoptotic gene expression by RTPCR. CM from co-cultured hWJMSCs and IL-2 NK cells inhibited NK and BC cell proliferation, increased expression of Bax and p53 and decreased the number of Bcl-2 in BC cells. In conclusion, CM of co-culture IL-2 treated NK cells and hWJMSCs induce apoptosis in BC cells as indicated by increased Bax and p53 expression and decreased Bcl-2 expression.

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Background: Breast cancer (BC) is the leading cause of death cancer in women. Cancer therapies using TNFα and IFNγ have been recently developed by direct effects and activation of immune responses. This study was performed to evaluate the effects of TNFα and IFNγ directly, and TNFα and IFNγ secreted by Conditioned Medium-human Wharton’s Jelly Mesenchymal Stem Cells (CM-hWJMSCs) toward apoptosis of BC cells (MCF7).Materials and Methods: BC cells were induced by TNFα and IFNγ in 175 and 350ng/mL, respectively. CM-hWJMSCs were produced by co-culture hWJMSCs and NK cells that secreted TNFα, IFNγ, perforin (Prf1), granzyme B (GzmB) for treating BC cells. The BC cells were treated with CM-hWJMSCs in 50%. The expression of apoptotic genes Bax, p53, and the antiapoptotic gene Bcl-2 were determined using RT-PCR.Results: TNFα and IFNγ at concentration of 350 ng/mL induced higher Bax expression compared to 175 ng/mL. TNFα and IFNγ 350 ng/mL, 175 ng/mL induced p53 expression, whilst TNFα and IFNγ at 350 ng/mL decreased Bcl-2 expression. Perf1, GzmB, TNFα and IFNγ-containing CM-hWJMSCs induced significantly apoptosis percentage, induced Bax expression, but did not effect p53, Bcl-2 expression.Conclusion: TNFα and IFNγ directly induce Bax, p53, decrease Bcl-2 gene expression. The Prf1, GzmB, TNFα, IFNγ-containing CM-hWJMSCs induce apoptosis and Bax expression.Keywords: breast cancer, Wharton’s Jelly mesenchymal stem cells, TNFα, IFNγ

Introduction:The most cases of cancer death, which are in the first rank among cancers suffered by women is breast cancer. The breast cancer therapy for patients has been done, but still not optimal, so it is necessary to understand the mechanism of therapy in model cell of breast cancer.Aim:This study aim to develop an isolation technique of breast cancer cell from patients as a cancer cell model.Material and methods:Breast cancer cell isolation is performed by enzymatic methods using the collagen I and hyaluronidase. Then, breast cancer cells were characterized using flow cytometry based on the CD44/CD24 expression where MDA-MB468 and MCF-7 breast cancer cell lines were used as positive controls. Estrogen receptor (ER), progesterone receptor (PR), p53, HER2, and Ki67 expression were assessed using an immunohistochemistry assay.Result and Discussion:The morphology of cancer cells was fibroblast like cells on the day 7th after isolation. Isolated breast cancer cells expressed 95.33±0.47% of CD44+/CD24+ and human epidermal growth factor receptor 2 (HER2) low expressions. Isolation of breast cancer cells can use In-house enzymatic protocol. Isolated breast cancer showed the same expression as MDA-MB468 (CD44+/CD24+) and HER2- compared to MCF-7 cell lines (CD44-/CD24+).Conclusion:These cells belonged to a basal type of breast carcinoma and expressed CD44+/CD24+, then isolated BCCs can be used as model cancer cells for further research.