Jong Bin Kim

The alkaloid Berberine inhibits the growth of anoikis-resistant MCF-7 and MDA-MB-231 breast cancer cell lines by inducing cell cycle arrest.

Berberine is a pure phenanthren alkaloid isolated from the roots and bark of herbal plants such as Berberis, Hydrastis canadensis and Coptis chinensis. Berberine has been established to inhibit the growth of breast cancer cells, but its effects on the drug resistance and anoikis-resistance of breast cancer cells have yet to be elucidated. Anoikis, or detachment-induced apoptosis, may prevent cancer progression and metastasis by blocking signals necessary for survival of localized cancer cells. Resistance to anoikis is regarded as a prerequisite for metastasis; however, little is known about the role of berberine in anoikis-resistance. We established anoikis-resistant cells from the breast cancer cell lines MCF-7 and MDA-MB-231 by culturing them on a Poly-Hema substratum. We then investigated the effects of berberine on the growth of these cells. The anoikis-resistant cells had a reduced growth rate and were more invasive than their respective adherent cell lines. The effect of berberine on growth was compared to that of doxorubicine, which is a drug commonly used to treat breast cancer, in both the adherent and anoikis-resistant cell lines. Berberine promoted the growth inhibition of anoikis-resistant cells to a greater extent than doxorubicine treatment. Treatment with berberine-induced cell cycle arrest at G0/G1 in the anoikis-resistant MCF-7 and MDA-MB-231 cells as compared to untreated control cells. In summary, these results revealed that berberine can efficiently inhibit growth by inducing cell cycle arrest in anoikis-resistant MCF-7 and MDA-MB-231 cells. Further analysis of these phenotypes is essential for understanding the effect of berberine on anoikis-resistant breast cancer cells, which would be relevant for the therapeutic targeting of breast cancer metastasis.

Berberine Diminishes the Side Population and ABCG2 Transporter Expression in MCF-7 Breast Cancer Cells

The plant alkaloid berberine has many biological activities including the ability to induce cell cycle arrest and apoptosis, making it a potentially useful agent for targeting cancer cells. We have analyzed the effects of berberine on MCF-7 breast cancer cells. Berberine was added to MCF-7 cells in culture, and proliferation, side population (SP) cells and expression of ABCG2 were examined. Berberine caused a dose-dependent reduction in proliferation. Hoechst 33,342 dye staining and FACS analysis revealed that berberine treatment caused a decrease in SP cells relative to untreated controls. In addition, berberine treatment was associated with a decrease in expression of ABCG2 relative to untreated controls. These results indicate that the growth inhibitory effects of berberine treatment on MCF-7 cells may be partly via effects on SP and ABCG2 expression. Further work is warranted to explore whether berberine may be a novel therapeutic drug useful for targeting breast cancer stem cells.

Berberine inhibits growth of the breast cancer cell lines MCF-7 and MDA-MB-231.

The effects of berberine on the behavior of breast tumors have not yet been established. To determine whether this compound is useful in the treatment of breast cancer, we analyzed the impact of berberine on the human breast cancer cell lines MCF-7 and MDA-MB-231 cells. Berberine was added to proliferating MCF-7 and MDA-MB-231 cells in culture. Following treatment, changes in cell growth characteristics such as proliferation, cell cycle duration, and the degree of apoptosis were assayed. Following berberine treatment, a time-dependent reduction in proliferation was observed in both cell lines at differing concentrations: 20 microM for MCF-7 and 10 microM for MDA-MB-231 cells. Annexin V staining showed an increase in apoptosis in both cell lines of 31 % in MCF-7 and 12 % in MDA-MB-231 cells compared to their respective controls. In addition, 12 % of the MCF-7 cells were arrested at G0/G1, compared to 62 % of control cells. These results demonstrate that treatment with berberine inhibits growth in both MDA-MB-231 and MCF-7 cells. In addition, they show that this partly occurs through the induction of apoptosis in MDA-MB-231 cells, and through both cell cycle arrest and induction of apoptosis in MCF-7 cells. Thus, berberine may be a novel therapeutic drug for breast cancer.

Berberine Inhibited the Growth of Thyroid Cancer Cell Lines 8505C and TPC1

Purpose Thyroid cancer is the most common malignancy in Korean females and can be treated with good prognosis. However, drugs to treat aggressive types of thyroid cancer such as poorly differentiated or anaplastic thyroid cancer have not yet been established. To that end, we analyzed the effects of berberine on human thyroid cancer cell lines to determine whether this compound is useful in the treatment of aggressive thyroid cancer. Materials and Methods The two thyroid cancer cell lines 8505C and TPC1, under adherent culture conditions, were treated with berberine and analyzed for changes in cell growth, cell cycle duration, and degree of apoptosis. Results Following berberine treatment, both cell lines showed a dose-dependent reduction in growth rate. 8505C cells showed significantly increased levels of apoptosis following berberine treatment, whereas TPC1 cells showed cell cycle arrest at the G0/G1 phase. Immunobloting of p-27 expression following berberine treatment showed that berberine induced a little up-regulation of p-27 in 8505c cells but relatively high up-regulation of p-27 in TPC1 cells. Conclusion These results suggest that berberine treatment of thyroid cancer can inhibit proliferation through apoptosis and/or cell cycle arrest. Thus, berberine may be a novel anticancer drug for the treatment of poorly differentiated or anaplastic thyroid cancer.

Sulforaphane Inhibits Oral Carcinoma Cell Migration and Invasion In Vitro

Sulforaphane is a predominant isothiocyanate in Brassica oleracea, a family of cruciferous vegetables, and is known to be inversely related to the risk of various types of human carcinomas. Studies using oral carcinoma cell lines are scarce, however, and the role of sulforaphane on oral carcinoma cell metastasis is yet to be determined. In this study, the growth inhibition of oral carcinoma cell lines by sulforaphane was determined using aqueous soluble tetrazolium salts, and the growth of various oral cancer cell lines was attenuated. The migration and invasion activities of the cells also decreased, as observed in monolayer scratch assays and transwell invasion experiments. The molecular change behind the impairment of the migration and invasion was investigated via secreted metalloprotease level detection using Multiplex protein analysis kits. At the molecular level, the secreted forms of MMP‐1 and MMP‐2 were down‐regulated. The expressions of MMP‐1 and MMP‐2 did not change when a conventional tumoricidal agent paclitaxel was used. These findings indicate that sulforaphane may have therapeutic potential as an inhibitor of metastasis in oral carcinoma patients. Copyright

Interleukin-8 is related to poor chemotherapeutic response and tumourigenicity in hepatocellular carcinoma

AIM Interleukin-8 (IL-8) has been suggested as a prognostic biomarker for human hepatocellular carcinoma (HCC), but its roles in HCC progression and drug resistance have not been studied. This study investigates the role and underlying mechanism of IL-8 in the chemoresistance and progressive growth of HCC. METHODS The change of chemosensitivity and proportion of side population in hepatoma cells was examined by cell growth and flow cytometric analyses after anti-cancer treatments or knockdown of IL-8. Expression of IL-8 and ATP-binding cassette (ABC) transporters in hepatoma cells, xenograft and clinical HCC tissues was determined by Western blot and immunohistochemical analyses. Tumourigenicity of hepatoma cells was evaluated in vivo after silencing IL-8 gene. RESULTS Treatment of hepatoma cells with anti-cancer drugs increased the production of IL-8 and its receptor, as well as the proportion of side population (SP). Exogenous IL-8 increased the SP fraction and expression of multidrug resistance-1, decreasing the drug sensitivity. Silencing of IL-8 gene decreased the ratio of SP cells and drug resistance properties. Both IL-8 and ABC transporters were highly expressed in xenograft and clinical HCC tissues, and knockdown of IL-8 significantly reduced tumour size in vivo. CONCLUSION Anti-cancer drug-induced IL-8 secretion increased the expression of ABC transporters and SP cells, promoting the growth of HCC in vitro. Thus IL-8 may be a potential therapeutic target in the treatment of HCC.

Entrapped doxorubicin nanoparticles for the treatment of metastatic anoikis-resistant cancer cells

Metastasized and chemoresistant secondary breast cancer treatment commonly shows very low efficacy. A new efficient treatment method is required to overcome the limitation against the secondary breast cancer. In this study, anoikis-resistant breast cancer cells, MDA-MB-231 and MCF-7 were developed as models of chemoresistant and metastatic breast cancer. Doxorubicin encapsulating human serum albumin nanoparticles (HSA+DOX NPs) were fabricated to confirm the benefits of nanoparticles at the treatment of anoikis-resistant breast cancer cells. The side population (SP) fraction in the anoikis-resistant cancer cells was higher than the parental cells. HSA+DOX NPs were more cytotoxic to anoikis-resistant cancer cells than free doxorubicin. The confocal microscope images demonstrated HSA+DOX NPs to deliver more doxorubicin into cells compared to the free doxorubicin by bypassing the drug efflux pump systems of anoikis-resistant cancer cells. In this study, a nanomedicine-based drug delivery carrier shows a potential in treating a metastasized and chemoresistant breast cancer.

CD24 cross-linking induces apoptosis in, and inhibits migration of, MCF-7 breast cancer cells

BackgroundThe biological effects of CD24 (FL-80) cross-linking on breast cancer cells have not yet been established. We examined the impact of CD24 cross-linking on human breast cancer cell line MCF-7.MethodsMCF-7 and MDA-MB-231 cells were treated with anti-rabbit polyclonal IgG or anti-human CD24 rabbit polyclonal antibodies to induce cross-linking, and then growth was studied. Changes in cell characteristics such as cell cycle modulation, cell death, survival in three-dimensional cultures, adhesion, and migration ability were assayed after CD24 cross-linking in MCF-7.ResultsExpression of CD24 was analyzed by flow cytometry in MDA-MB-231 and MCF-7 cells where 2% and 66% expression frequencies were observed, respectively. CD24 cross-linking resulted in time-dependent proliferation reduction in MCF-7 cells, but no reduction in MDA-MB-231 cells. MCF-7 cell survival was reduced by 15% in three-dimensional culture after CD24 cross-linking. Increased MCF-7 cell apoptosis was observed after CD24 cross-linking, but no cell cycle arrest was observed in that condition. The migration capacity of MCF-7 cells was diminished by 30% after CD24 cross-linking.ConclusionOur results showed that CD24 cross-linking induced apoptosis and inhibited migration in MCF-7 breast cancer cells. We conclude that CD24 may be considered as a novel therapeutic target for breast cancer.

The CD49d+/high subpopulation from isolated human breast sarcoma spheres possesses tumor-initiating ability.

Primary breast sarcomas (PBSs) that arise from mammary stroma are very rare, highly aggressive and therapy- resistant tumors with a heterogeneous phenotype. In this study, we sought to identify tumor-initiating cells (TICs) in PBSs and to describe their features. We isolated long-term self-renewing sarcospheres (designated NDY-1) from primary breast carcinosarcoma tissue (sarcoma component >95%) using the anchorage-independent culture method. NDY-1 spheres expressed various mesenchymal cell markers, and their tumorigenic potential was markedly reduced in adherent culture conditions, compared to spheres. Screening for integrins revealed a marked decrease in CD49d expression in adherent culture conditions of NDY-1. The CD49d+/high subpopulation sorted from NDY-1 spheres displayed higher cell viability and sphere-forming ability than CD49d-/low population in vitro. Moreover, the CD49d+/high population displayed high tumor initiating ability in limiting dilution transplantation to NOD/SCID mice, and the xenotransplanted CD49d+/high population recapitulated the complexity of the original primary tumors. Greater doxorubicin resistance was exhibited by the CD49d+/high population, compared with the CD49d-/low population. Thus, our results collectively demonstrate that CD49d+/high cells from sarcospheres display enhanced sphere-forming, drug resistance and tumor-initiating abilities. To our knowledge, this is the first study to identify TICs from breast sarcoma.

Berberine-induced growth inhibition of epithelial ovarian carcinoma cell lines

Aim:  The aim of this study is to analyze the impact of berberine on the two human epithelial ovarian carcinoma cell lines OVCAR‐3 and SKOV‐3 in relation to the potential usefulness of berberine in the treatment of epithelial ovarian cancer.