Aktivitas Sitotoksik dan Antiproliferasi Fraksi n-Heksan Biji Al-pukat (Percea americana Mill.) Terhadap sel T47D

Avocado plants are widely used as traditional medicine in Indonesia. This research aimed to determine the cytotoxic and antiproliferation activity of avocado seed extract and fractions on the T47D breast cancer cell line. Extraction was done by maceration method using 96% ethanol, then fractionation was done by liquid-liquid partition using n-hexane, chloroform, and ethyl acetate. Cytotoxic and antiproliferation tests were done by the MTT method. Apoptosis activity was investigated by the double staining method, using acridine orange-propidium iodide as a staining reagent. Results showed that the n-hexane fraction of avocado seed had a cytotoxic activity with IC50 27.9 µg/mL. N-hexane fraction of avocado seed inhibited proliferation and induced apoptosis of T47D cell line.

The comparison of Zn(II) arginine dithiocarbamate cytotoxicity in T47D breast cancer and fibroblast cells

BACKGROUND: With essential metals being studied and developed as anticancer agents, this study aims to explore the anticancer activity of Zn(II) arginine dithiocarbamate in the T47D and fibroblast cell lines. METHOD: The Zn(II) arginine dithiocarbamate complex was prepared by the in situ method and characterized using infra-red spectroscopy, melting point, X-ray fluorescence, and X-ray diffraction instruments. The complex compound was tested for its cytotoxicity to the T47D breast cancer and fibroblast cell lines. RESULTS: The cytotoxicity of the Zn(II) arginine dithiocarbamate complex to the T47D breast cancer cell line obtained IC50 = 3.16 μg/mL, while cisplatin obtained IC50 = 28.18 μg/mL. The cytotoxicity of the Zn(II) arginine dithiocarbamate complex to fibroblast cells obtained IC50 = 8709.63 μg/mL. CONCLUSION: The Zn(II) arginine dithiocarbamate complex has increased active cytotoxicity compared to cisplatin in inducing morphological changes in the T47D breast cancer cell line and is relatively non-toxic to fibroblast cells.

Characterization of SN38-Resistant T47D Breast Cancer Cell Sublines Overexpressing BCRP, MRP1, MRP2, MRP3, and MRP4

BackgroundAlthough several novel resistant breast cancer cell lines have been established, only a few resistant breast cancer cell lines overexpress breast cancer resistance proteins (BCRP). The aim of this study was to establish new resistant breast cancer cell lines overexpressing BCRP using SN38 (7-ethyl-10-hydroxycamptothecin), an active metabolite of irinotecan and was to discover genes and mechanisms associated with multidrug resistance.MethodsSN38-resistant T47D breast cancer cell sublines were selected from the wild-type T47D cells by gradually increasing SN38 concentration. The sensitivity of the cells to anti-cancer drugs was assessed by 3-(4,5-methylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. Expression profiles of the resistance-related transporters were examined using the resistance diagnostic kit (Drugsporter®), real time RT-PCR, and western blot analysis. Intracellular drug accumulation in the resistant cells was determined using flow cytometry.ResultsThe SN38-resistant T47D breast cancer cell sublines T47D/SN120 and T47D/SN150 were established after long-term exposure (more than 16 months) of wild-type T47D cells to 120 nM and 150 nM SN38, respectively. T47D/SN120 and T47D/SN150 cells were more resistant to SN38 (14.5 and 59.1 times, respectively), irinotecan (1.5 and 3.7 times, respectively), and topotecan (4.9 and 12 times, respectively), than the wild-type parental cells. Both T47D/SN120 and T47D/SN150 sublines were cross-resistant to various anti-cancer drugs. These resistant sublines overexpressed mRNAs of MRP1, MRP2, MRP3, MRP4, and BCRP. The DNA methylase inhibitor 5-aza-2'-deoxycytidine and the histone deacetylase inhibitor trichostatin A increased the expression levels of BCRP, MRP1, MRP2, MRP3, and MRP4 transcripts in T47D/WT cells. Drug accumulation was found to be lower in T47D/SN120 and T47D/SN150 cells, compared to that in T47D/WT cells. However, treatment with known chemosensitizers increased the intracellular drug accumulation and sensitivity of anti-tumor agents.ConclusionT47D/SN120 and T47D/SN150 cells overexpressed MRP1, MRP2, MRP3, MRP4, and BCRP due to the suppression of epigenetic gene silencing via DNA hypermethylation and histone deacetylation. Although these resistant cells present a higher resistance to various anti-cancer drugs than their parental wild-type cells, multidrug resistance was overcome by treatment with chemosensitizers. These SN38 resistant T47D breast cancer cell sublines expressing resistance proteins can be useful for the development of new chemosensitizers.

Dexamethasone Reduces Cell Adhesion and Migration of T47D Breast Cancer Cell Line

Background: Aberrant expression of cell adhesion molecules and matrix metalloproteinase (MMPs) plays a pivotal role in tumor biological processes including progression and metastasis of cancer cells. Targeting these processes and detailed understanding of their underlying molecular mechanism is an essential step in cancer treatment. Dexamethasone (Dex) is a type of synthetic corticosteroid hormone used as adjuvant therapy in combination with current cancer treatments such as chemotherapy in order to alleviate its side effects like acute nausea and vomiting. Recent evidences have suggested that Dex may have antitumor characteristics. Objective: Dex affects the migration and adhesion of T47D breast cancer cells as well as cell adhesion molecules e.g., cadherin and integrin, and MMPs by regulating the expression levels of associated genes. Methods: In this study, we evaluated the cytotoxicity of Dex on the T47D breast cancer cell line through MTT assay. Cell adhesion assay and wound healing assay were performed to determine the impact of Dex on cell adhesion and cell migration, respectively. Moreover, real-time PCR was used to measure the levels of α and β integrin, E-cadherin, N-cadherin, MMP-2, and MMP-9. Results: Dex decreased the viability of T47D cells in a time and dose-dependent manner. Cell adhesion and migration of T47D cells were reduced upon Dex treatment. The expression of α and β integrin, E-cadherin, N-cadherin, MMP-2, and MMP-9 were altered in response to the Dex treatment. Conclusion: Our findings demonstrated that Dex may have a role in the prevention of metastasis in this cell line.

The Cytotoxic Activity of Annona muricata Linn Leaves Ethanolic Extract (AMEE) on T47D Breast Cancer Cell Line

Breast cancer is the most common cancer in women throughout the world, with new cases and deaths which continue to increase. Soursop leaves (Annona muricata L) have been used extensively in traditional medicine, including cancer. Acetogenin, alkaloids, and phenols contained in soursop leaves are known to have anti-cancer effects. Among them, acetogenin has the most dominant role and reported to have a cytotoxic effect on various cancer cell lines. This study aims to determine the cytotoxic activity of soursop leaf ethanol extract on T47D breast cancer cell line. Measurement of cytotoxic activity was carried out by the MTT method, and the viability percentage of T47D cells was calculated based on the absorbance values in the treatment, cell control, and media control groups of each replicate. The correlation between extract concentration and viability percentage of the T47D cell line was outlined in the regression equation to obtain the IC50 value. IC50 values of 109.91 ± 3.04 with R values 0.975 and R2 0.9508 obtained. R values close to 1 indicated a strong correlation between extract concentration and the percentage of living T47D cells. Meanwhile, the amount of R2 suggested that the level of AMEE had a 95.08% influence on the rate of cell viability, and the other 4.92% influenced by factors other than the AMEE dose. These results indicated that the ethanol extract of soursop leaves has a cytotoxic effect and has the potential to inhibit T47D cell proliferation in vitro.

Uji sitotoksik metformin hidroklorida terhadap sel kanker payudara T47D

Abstrak. Latar Belakang. Kanker payudara merupakan penyakit berlebihnya pertumbuhan atau tidak terkendalinya perkembangan sel kanker payudara. Kanker merupakan suatu penyakit yang disebabkan kelainan genetik berupa mutasi DNA yang menyebakan hilangnya kontrol pertumbuhan. Gangguan genetik ini menyebabkan terganggunya siklus sel dan apoptosis. Metformin merupakan suatu antihiperglikemik yang digunakan pada pasien diabetes melitus tipe 2. Penurunan risiko kanker terjadi pada pasien diabetes melitus tipe 2 yang menggunakan metformin. Uji sitotoksik untuk agen anti kanker merupakan uji skrining awal untuk menilai potensi efek anti kanker. Tujuan penelitian ini adalah untuk mengetahui efek sitotoksik metformin hidroklorida terhadap pertumbuhan sel kanker payudara T47D. Metode: Penelitian ini merupakan penelitian eksperimental uji invitro terhadap sel kanker payudara T47D yang dipaparkan metformin HCl konsentrasi 5000; 2500; 1250; 312.5 dan 156,25 μM selama 24 jam. Sebagai pembanding digunakan paclitaxel konsentrasi 1000; 500; 250; 31,25 dan 15,625 nM. Uji sitotoksik menggunakan metode MTT untuk menentukan IC50.Data dianalaisis menggunakan analisa probit. Hasil : IC50 metformin HCl adalah 13457.3 ± 1096,5 μM. IC50 paclitaxel adalah 1577.2 ± 115.3 nM. Efek anti kanker metformin lebih kecil dibanding paclitaxel. Kata Kunci: Metformin HCl, T47D, uji sitotoksik, IC50 Abstract. Breast cancer is a disease in which there is excessive growth or uncontrolled development of breast tissue cells. Cancer is a disease caused by genetic disorders caused by DNA mutations that cause loss of growth control. This genetic disorder affects the cell cycle and cell apoptosis and causes the formation of cancer. Metformin is an antihyperglycemic in type 2 diabetes mellitus patient. The decrease in cancer risk occured in patients with type 2 diabetes mellitus who used metformin. Cytotoxic test for agent anti cancer is screening test to investigate the potency cancer effect of substance. The goal of this study was determining the cytotoxic effect of metformin hydrochloride to T47D breast cancer cell. The method : This sudy was experimental study, invitro test to T47D breast cancer cell using metformin HCl 5000; 2500; 1250; 312.5; and 156.25 μM for 24 hours. Paclitaxel used as postiive control with concentration were 1000; 500; 250; 31,25 and 15,625 nM. Cytotoxic test using MTT method to determine IC50. Data were analyzed using probit analysis using SPSS 22 version. The result of cytotoxic test showed that IC50 metformin HCl was 13457.3 ± 1096,5 μM. While IC50 paclitaxel as control was 1577.2 ± 115.3 nM. The effect of cancer metformin HCl was lower than paclitaxel.Keywords: Metformin HCl, T47D, cytotoxic test, IC50

UJI SITOTOKSIK METFORMIN HIDROKLORIDA TERHADAP SEL KANKER PAYUDARA T47D

Abstrak. Latar Belakang. Kanker payudara merupakan penyakit berlebihnya pertumbuhan atau tidak terkendalinya perkembangan sel kanker payudara. Kanker merupakan suatu penyakit yang disebabkan kelainan genetik berupa mutasi DNA yang menyebakan hilangnya kontrol pertumbuhan. Gangguan genetik ini menyebabkan terganggunya siklus sel dan apoptosis. Metformin merupakan suatu antihiperglikemik yang digunakan pada pasien diabetes melitus tipe 2. Penurunan risiko kanker terjadi pada pasien diabetes melitus tipe 2 yang menggunakan metformin. Uji sitotoksik untuk agen anti kanker merupakan uji skrining awal untuk menilai potensi efek anti kanker. Tujuan penelitian ini adalah untuk mengetahui efek sitotoksik metformin hidroklorida terhadap pertumbuhan sel kanker payudara T47D. Metode: Penelitian ini merupakan penelitian eksperimental uji invitro terhadap sel kanker payudara T47D yang dipaparkan metformin HCl konsentrasi 5000; 2500; 1250; 312.5 dan 156,25 μM selama 24 jam. Sebagai pembanding digunakan paclitaxel konsentrasi 1000; 500; 250; 31,25 dan 15,625 nM. Uji sitotoksik menggunakan metode MTT untuk menentukan IC50.Data dianalaisis menggunakan analisa probit. Hasil : IC50 metformin HCl adalah 13457.3 ± 1096,5 μM. IC50 paclitaxel adalah 1577.2 ± 115.3 nM. Efek anti kanker metformin lebih kecil dibanding paclitaxel. Kata Kunci: Metformin HCl, T47D, uji sitotoksik, IC50 Abstract. Breast cancer is a disease in which there is excessive growth or uncontrolled development of breast tissue cells. Cancer is a disease caused by genetic disorders caused by DNA mutations that cause loss of growth control. This genetic disorder affects the cell cycle and cell apoptosis and causes the formation of cancer. Metformin is an antihyperglycemic in type 2 diabetes mellitus patient. The decrease in cancer risk occured in patients with type 2 diabetes mellitus who used metformin. Cytotoxic test for agent anti cancer is screening test to investigate the potency cancer effect of substance. The goal of this study was determining the cytotoxic effect of metformin hydrochloride to T47D breast cancer cell. The method : This sudy was experimental study, invitro test to T47D breast cancer cell using metformin HCl 5000; 2500; 1250; 312.5; and 156.25 μM for 24 hours. Paclitaxel used as postiive control with concentration were 1000; 500; 250; 31,25 and 15,625 nM. Cytotoxic test using MTT method to determine IC50. Data were analyzed using probit analysis using SPSS 22 version. The result of cytotoxic test showed that IC50 metformin HCl was 13457.3 ± 1096,5 μM. While IC50 paclitaxel as control was 1577.2 ± 115.3 nM. The effect of cancer metformin HCl was lower than paclitaxel. Keywords: Metformin HCl, T47D, cytotoxic test, IC50