Anti-migration Effect of Aaptos suberitoides Fraction in HCT-116 Colorectal Cancer Cell Line

Background: Colorectal cancer is the second leading cause of mortality and the most prevalent cancer worldwide. Most patients, who come with late-stage, have ineffective treatments and some side effects in chemotherapy. Aaptos suberitoides has potential anti-cancer effects due to its bioactive compounds such as aptamine. This study aimed to evaluate the migration inhibition effect of Aaptos suberitoides fraction in HCT-116 cell line.Methods: This study was an experimental study. Aaptos suberitoides specimen was taken in Tinjil Island and fractionated with ethyl acetate. HCT-116 cell line was added with Aaptos suberitoides fraction and cellular migration activity was observed in 48 hours of which the scratch assay was performed. The gap closure area was determined with ImageJ software.Results: The data showed that a low concentration of Aaptos suberitoides fraction inhibited migration activity in HCT-116 cell line as follow; 1 and 5 mg/L Aaptos suberitoides fraction inhibit 3-4 % cancer cell migration in 24 hours, and 10-11% inhibition in 48 hours, respectively. However, 10 mg/L fraction concentration only inhibited 7-14% of the migration effect.Conclusion: Aaptos suberitoides fraction suggests insignificant migration inhibition in colorectal cancer cells and only inhibits less than 15 % HCT-116 cell line

SENYAWA TRITERPENOID 3β-HIDROKSI-TIRUKAL-7-EN DARI EKSTRAK DAUN KAPI NANGO (Dysoxylum arborescens) DAN AKTIVITAS SITOTOKSIKNYA TERHADAP SEL KANKER PAYUDARA MCF-7

Senyawa triterpenoid 3β-hidroksi-tirukal-7-en (1) telah diisolasi dari ekstrak daun kapi nango (Dysoxylum arborescens). Struktur kimia senyawa ditentukan menggunakan data spektroskopi yang meliputi IR, 1H-NMR, 13C-NMR, MS dan perbandingan dengan senyawa yang telah dilaporkan sebelumnya. Aktivitas sitotoksik senyawa 1 terhadap sel kanker payudara MCF-7  menunjukkan nilai IC50 155,1 ppm

THE N-HEXANE FRACTION OF MYRMECODIA PENDANS INHIBITS CELL SURVIVAL AND PROLIFERATION IN COLON CANCER CELL LINE

Objective: Despite advanced treatment options available for colorectal cancer, many reported resistance and unresponsiveness to conventional chemotherapeutic agents. Therefore, it is urgent to discover a novel drug for colon cancer. Sarang Semut (Myrmecodia pendans), an Indonesian native plant, has been studied extensively due to its anti-cancer profiles. This study aimed to evaluate the anti-tumour activity of Sarang Semut in colon cancer cells.Methods: We evaluated cytotoxic activity of methanol extract as well as n-hexane and ethyl acetate fraction towards colon cancer cell lines (Caco-2 and HCT-116 cells) utilizing 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay. The most potent fraction was evaluated further in inhibiting cell survival using MTT assay and cell proliferation using trypan blue exclusion assay as well as a clonogenic assay.Results: Our data showed that the n-hexane fraction of Sarang Semut induces more cell death than the methanol extract and ethyl acetate fraction. Therefore, we analyzed the n-hexane fraction further and found that the inhibitory concentration 50% (IC50) of the n-hexane fraction was 24 and 30 parts per million (ppm) for Caco-2 and HCT-116 cells, respectively. Moreover, it inhibited cell growth as well as cell colony formation, in particular, shown by the plating efficiency (P<0.05) and colony area per seed (P<0.01) of the control group were different to the treatment group.Conclusion: The n-hexane fraction of Sarang Semut demonstrates a high potential antitumor activity in colon cancer cell line

The usage of antibiotics for prevention of contamination in 3T3-L1 preadipocyte culture

Background: 3T3-L1 preadipocyte is the most commonly used cell line in in vivo studies of obesity. One of the main concerns in 3T3-L1 preadipocyte culture is microorganism contaminations. The objective of this study was to determine the appropriate antibiotics to prevent contaminations in 3T3-L1 cultures. Method: This study used descriptive analysis. Frozen 3T3-L1 preadipocytes were thawed and cultured in DMEM-10% FBS-1% penicillin-streptomycin, DMEM-10% FBS-1% penicillin-streptomycin-fungiezone, or DMEM-10% FBS-0.2% ciprofloxacin 200 mg/100 ml. After 24-hour incubation, the cells were observed under the microscope for any change in the medium colour, presence of abnormal structures, and abnormality in cell morphology. Results: The usage of 1% penicillin-streptomycin, 1% penicillin-streptomycin-fungiezone, or 0.2% ciprofloxacin 200 mg/100 ml maintained the clean medium and conserved normal fibroblast-like morphology of the cells. Conclusion: This study suggested that 1% penicillin-streptomycin, 1% penicillin-streptomycin-fungiezone, or 0.2% ciprofloxacin 200 mg/100 ml can be utilized in 3T3-L1 preadipocyte cultures to prevent contaminations

Inhibition Capacity of the n-Hexane Fraction of Myrmecodia pendens as a Potential Anti-Cancer in Breast and Cervical Cancer: In Vitro Study

Breast cancer (BC) and cervical cancer (CC) have a high prevalence and mortality rate worldwide. Despite the availability of advanced treatment, resistance to conventional chemotherapies has emerged. Myrmecodia pendens, one of the species of Sarang Semut (local name), possess a potential of antitumor effects by inducing cell death different cancer cell entities. This study aimed to assess anti-tumor activities of n-hexane fraction of M. pendens in inhibiting cell survival and cell migration in BC and CC cells. M. pendens was extracted in methanol then fractionated using n-hexane or ethyl acetate. BC cells including MCF-7 (luminal A), HCC-1954 (HER2+) cells and CC Hela cells were treated with M. pendens extracts to evaluate cytotoxic activity using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay as well as anti-cell migration using scratch assay. We also analyzed inhibitory concentration 50 (IC50) of n-hexane fraction in BC and CC cells. We started with comparing cytotoxicity activities of methanol extract, ethyl acetate and n-hexane fractions of M. pendens. Data showed that the n-hexane fraction was the most potent inducing BC cell death. Therefore, we used the n-hexane fraction for further experiments. Interestingly, IC50 of this fraction in HCC-1954 and Hela cells were lower than in MCF-7 cells, 16; 13 and 60 ppm, respectively. Moreover, the low concentrations of n-hexane fraction inhibited HeLa cells migration, compared to control group (p<0.05). The n-hexane fraction of M. pendens shows promising anti-cancer agent, by inhibiting BC and CC cell survival as well as inhibiting CC cells migration.Keywords: breast cancer, cervical cancer, MTT assay, Sarang Semut, scratch assa

Seeding cell number required for optimal lipid accumulation during adipocyte differentiation using 3T3-L1 cell line

Obesity is one of the major causes of metabolic diseases such as diabetes and heart attack, and, hence, can lead to low quality of life. Elaborating adipocyte differentiation is very crucial for formulating the treatment and prevention of obesity. The objective of this study is to investigate the seeding cell number required to obtain optimum lipid accumulation during adipocyte differentiation using the 3T3-L1 cell line. Two sets of 5.48×104 (for Day 0 and Day 8 of differentiation), of 10.96×104 (for Day 8) and of 21.92×104 (for Day 8) of 3T3-L1 cells were seeded in each wells of a 12-well plate. Isobuthylmethylxanthine (IBMX), Dexamethasone, and Insulin-containing differentiation cocktails was added into the medium at Day 0 for 48 hours. The medium was changed every two days. Day 0 and Day 8 samples were then stained using Oil Red O and were examined under the microscope to observe the lipid droplets (red-coloured). The lipid droplets were quantified by measuring the absorbance at wavelength of 550 nm. In the study, seeding the number of 10.96×104 cells produced very significantly higher lipid accumulation, as compared with seeding the number of 5.48×104 cells. However, doubling the seeding number into 21.92×104 cells did not increase the lipid droplets significantly. This study found that the optimum seeding number to obtain the maximum lipid droplets during 3T3-L1 adipocyte differentiation was 10.96×104 cells

Anti-tumor activity of metformin in human epidermal growth factor receptor 2 positive breast cancer cells

Breast Cancer (BC) is the leading cause of cancer death in women. One BC subtype is very aggressive with amplification of human epidermal growth factor receptor 2 (HER2) protein. Although specific HER2+ targeting agents are available, most of HER2+ BC patients develop resistant to these agents. Recent studies show that metformin, is able to become anti-tumor in various cancer cells. This research aims to evaluate anti-tumor activities of metformin to HER2+ BC cells in both sensitive and resistant to trastuzumab. A series of assays were performed to evaluate metformin anti-tumor activities in HCC-1954 and SKBR-3 HER2+ BC cells. MTT assay was performed to evaluate cell death, and inhibitory concentration (IC50), while scratch assay was performed to assess inhibition of cell migration and clonogenic assay to assess cell proliferation. p<0.05 was considered to be significant. Metformin could suppress the number of HER2+ BC cells. Viability assay showed suppression of viable cells after metformin incubation of 60 and 600 µM compared to control, 30 and 90%, respectively. Surprisingly, IC50 of metformin was smaller in HER2+ BC HCC-1954 cells that resistant to trastuzumab compare to the sensitive one (SKBR-3). Both were below 1 µM, with R2 more than 0.95. Additionally, clonogenic assay showed less colony number and colony area with at least p < 0.05 in colony number and p < 0.01 in the area. In addition, metformin inhibited cell migration of HER2+ BC cells. Metformin shows a potency as anti-tumor by inducing cell death, inhibiting cell proliferation and cell migration of HER2+ BC cells