Mal J Nutr 20(1): 113 - 119, 2014

B-carotene Roles in Proliferation and Differentiation, Connexin and /3-casein Gene Expression of Mammary Gland Cells Line
Roosita K1, Rimbawan SW, Djuwita I2, Damanik MR1, Kusharto CMi, Damayanthi E1 & Nomura N3

ABSTRACT

Introduction: The objectives of this study were to examine the effect of 0-carotene on cells proliferation and differentiation, connexin (Cx43) and $-casein (Csn2) gene expression of mammary gland cells line (HC11). Cx43 involves development of gap junction intercellular communication, while Csn2 induces milk synthesis in differentiated mammary gland cells. Method: The HC11 cells were grown in medium RPMI 1640 supplemented with 10% heat-inactivated Fetal Bovine Serum (FBS), 50 mg/ml streptomycin and enhanced by epidermal growth factor (EGF), insulin, hydrocortisone and prolactin. Final concentration of 0-carotene in medium was 0.5, 1.5 and 5.0 AM respectively. Cell proliferation was determined by a colorimetric assay for assessing cell viability (MTT assays). Gene expression was analysed by reverse trancriptation polymerase chain reaction (RT-PCR) and gel electrophoresis. Results: Triton X-114 as a negative control siginificanly decreased proliferation of HC11 cell line as compared to its control (<0.05%). Meanwhile, 0-carotene even in supraphysiologycal concentration [5).N.] did not affect proliferation of HC11 cell line. The structure of mammosphere of HC11 cell line treated with supraphysiological concentration 0-carotene [5.0 04] was largest with higher density and tightly connected with their neighbouring cells. Cx43 and Csn2 saw higher expression on 0-carotene treated HC11 cell lines. Conclusion: 0-carotene did not inhibit cell proliferation; meanwhile, differen¬tiation of mammary gland cells line (HC11) represented by mammosphere development, and genes expression of connexin (Cx43) and J3-casein (Csn2) were induced by 0-carotene.

Keywords: 0-carotene, fi-casein (Csn2), connexin (Cx43), genes expression, lactogenesis mammary gland cells line (HC11), mammosphere development

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