Combination Curcuma longa and Phyllanthus niruri Extract Potentiate Antiproliferative in Triple Negative Breast Cancer MDAMB-231 Cells.

BACKGROUND: Triple negative breast cancer cells (TNBC) are a small part of cancer-inducing cells in breast cancer, which are characterized by high metastatic and self-renewal. Self-renewal has the ability to renew itself and loses control of proliferation. Curcuma longa extract (CL) and Phyllanthus niruri extract (PN) known to have anti-proliferative effects on cancer cells. However, the effects of combination CL and PN on TNBC proliferation still unclear. AIMS: This study aimed to evaluate the antiproliferative effects of the combination CL and PN on TNBC MDAMB-231 and attempted to elucidate the underlying molecular mechanisms. SUBJECTS AND METHODS: The dried rhizomes of Curcuma longa and the herbs of Phyllanthus niruri were macerated with ethanol for 72 h.The antiproliferative and synergistic effects of combination CL and PN were investigated using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay. Combination index values were calculated using CompuSyn (ComboSyn, Inc, Paramus, NJ). The cell cycle and apoptosis assay were determined by propidium iodide (PI) and PI-AnnexinV assay under flow cytometer, respectively. The intracellular ROS levels were evaluated using 2',7'-Dichlorodihydrofluorescein diacetate (DCFDA) assay. The mRNA expressions of proliferation-related genes in the cells were determined using bioinformatic assay. RESULTS: The CL and PN single treatment caused a potent and dose-dependent decrease in the percentage of viable cells with IC50 value of 13 µg/mL and 45 µg/mL for 24 h, respectively. The combination index values of the different combinations ranged from 0.08 - 0.90, indicating slightly strong to very strong synergistic effects. The combination of CL and PN also remarkably induced the S- and G2/M-phases cell cycle arrest that leading to apoptosis induction. Furthermore, the combination of CL and PN treatment induced the intracellular reactive oxygen species (ROS) levels. Mechanistically, the AKT1, EP300, STAT3 and EGFR signaling as potential targets of combination CL and PN in antiproliferation and antimetastatic of TNBC. CONCLUSIONS: The combination of CL and PN exerted promising antiproliferative effects in TNBC. Therefore, CL and PN may be considered a potential source for the development of potent anticancer drugs for breast cancer treatment.

The PD-L1 Expression Among Triple-Negative Breast Cancer Patients in Universitas Sumatera Utara Teaching Hospital, Indonesia.

Background: The emerging role of precision medicine among in oncologic science is a potentially explorable area to solve long-standing implication of triple-negative breast cancer (TNBC)'s management, especially by identifying programmed cell death ligand-1 (PD-L1) in its population. Objective: To describe the PD-L1's expression among TNBC populations in our Indonesia-based centers. Methods: This descriptive study was conducted in the teaching hospitals of Universitas Sumatera Utara ranging from April 2019 to July 2020. Our investigation encompassed female individuals with histopathologically confirmed TNBC and complete medical record data, especially the status of PD-L1 expression to be reported in this study. We use Daco 22C3 antibodies to confirm the latter protein immunohistochemical positivity after thorough specimen preparation and staining. Results: This study included 60 females with TNBC of which 40 participants were issued in the final report. Our populations were dominated by middle-aged individuals (41-50 years old; 45%), and remarkable presentation of LVSI, angioinvasion, or lymph-node metastatic status. In PD-L1 expression status, our study reported that 45.0% patients were confirmed with confirmed PD-L1 positivity. Conclusion: PD-L1 expression among TNBC patients is concordant with the global report hence anti-PD-1-based treatment trial in Asia or even Indonesia region should be considered.

Regulation of p53 and survivin by Curcuma longa extract to caspase-3 dependent apoptosis in triple negative breast cancer cells.

Aim Triple negative breast cancer cells (TNBC) are the population of breast cancer cells that are responsible for cancer recurrence and apoptosis resistance. Unfortunately, current therapies have limited efficacy to TNBC population due to apoptosis resistance and chemoresistance. Tumour suppressor p53 and survivin are primary targets for TNBC therapy. Consequently, a search for a natural compound which targets p53 and survivin is needed to further advance TNBC treatment. Curcuma longa extract (CL), a natural compound induces apoptosis in several cancer cells by targeting various molecules and possess fewer side effects. However, a possible potential of CL as p53- and survivin modulating agent in TNBC cells has not been investigated. Methods MDAMB-231 cells were treated with several concentration of CL, after which, viability, p53 gene expression, surviving protein expression, and caspase-3 protein expression were evaluated. Results After 24-h treatment, CL possessed cytotoxic effect with IC50 value of 13 µg/mL. Treatment with 1.625, 3.25, 6.5, and 13 µg/mL of CL resulted in 2.70-25.80% increase in caspase-3 expression levels followed by 94.60 - 21.60% decrease in survivin protein levels. CL induced remarkably p53 gene expression ratio up to 5-fold at 13 µg/mL. Survivin protein levels were inversely proportional to p53 accumulation levels. Low survivin protein levels combined with high levels of p53 accumulation were correlated to higher apoptotic rates. Conclusion p53 and survivin as molecular targets of CL contribute to caspase-3-dependent apoptosis in TNBC cells and this compound represents an attractive p53- and survivin modulating agent in TNBC.

Role of Hormonal Receptor in Predicting Sentinel Lymph Node Metastasis in Early Breast Cancer.

Background: Sentinel lymph node biopsy is one of the minimally invasive techniques that can confirm the presence of metastasis of regional lymph nodes in cancer. Sentinel lymph node biopsy can be done with a lymph mapping technique using blue-dye, radiotracer, or a combination of both. In developing countries, sentinel lymph node biopsy is often done with a single agent, which is the blue dye. The limitation of conducting SLNB in Indonesia is the availability of patent blue dye and radioisotope tracer. To overcome that, a hormonal receptor is expected to aid in predicting sentinel lymph node metastasis. Objective: The aim of this study was to investigate the hormonal receptor as a prognostic factor of sentinel lymph node metastasis in breast cancer. Methods: This study was conducted in Universitas Sumatera Utara Teaching Hospital with the acknowledgment from the Ethics Committee of the respected hospital by the number of 116/KEP/USU/2020. Total of 51 patients participated in this research. Results: Statistically, the p-value in each immunohistochemistry group is > 0.05 in all ER (+) / PR (+); ER (+) / PR (-); ER (-) / PR (+) groups. This shows that there is no significant relationship between hormonal receptors on sentinel lymph node metastases. Conclusion: The statistical evaluation showed that there is no significant correlation between the hormonal receptor and sentinel lymph node metastasis (p>0.05), but is found clinically significant. Therefore, hormonal receptors should be considered as a predicting factor for sentinel lymph node metastasis.

Accuracy of Methylene Blue Test as Single Technique for Sentinel Lymph Node Biopsy in Early Stages Breast Cancer.

AIM: Sentinel Lymph Node Biopsy (SLNB) establishes as a gold standard for diagnostic lymph node involvement in early breast cancer. Most of the developed country does not have radiotracer and nuclear medicine facilities. Unless in Indonesia there is Methylene Blue as an alternative agent for SLNB. This study measure accuracy of sentinel lymph node biopsy as a single technique using the Methylene Blue test. METHODS: This cross-sectional study enrolled 60 female patients with breast cancer stage I-II. We performed SNB using 2-5 cc of 1% Methylene-blue dye (MBD) injected to periareolar tissue and proceeded with axillary lymph nodes dissection (ALND). The histopathology results of sentinel nodes (SNs) and axillary lymph nodes (ALNs) analyze for diagnostic value assessments. RESULTS: The identification rate of SN was 97.62 %, and the median number of identified SNs was 4 (2-7). Sentinel node metastasis was found in (19/60) % cases and % of them were macrometastases. The sensitivity and specificity of MBD were 91.67% and 96.67% respectively. The negative predictive value (NPV) of SNs to predict axillary metastasis was 96.67% (95% CI, 81-99%). CONCLUSION: Injection of 1% MBD as a single technique in breast cancer SNB has a favorable identification rate and predictive value.

Mesenchymal stem cells under hypoxia condition inhibit peritoneal adhesion by suppressing the prolonged release of interleukin-6.

Aim To investigate the role of hypoxia-preconditioned mesenchymal stem cells (H-MSCs) in preventing peritoneal adhesion by regulating IL-6 at days 6 and 12. Methods Twenty-four PAs rat model weighing 250 g to 300 g were randomly allocated into 4 groups: sham (Sh), control (C), H-MSCs treatment group at dose 1.5 x 106(T1) and 3 x 106(T2). To induce H-MSCs, all MSCs population were incubated under hypoxia state (5% O2 ), 5% CO2, and 37oC for 24 hours. Expression level of IL-6 was performed using ELISA. Morphological appearance of adhesion was observed by visualizing the existence of adhesion formation in intestinal. Results In this study we found that there was a trend of decrease of IL-6 level on day 6 following MSCs treatments. Interestingly, there was a significant decrease of IL-6 level on day 12 in all treatment groups. Also, no adhesion occurred in T2 group. Conclusions H-MSCs prevent PA development by suppressing the prolonged release of IL-6 at proliferation phase.