|Title : Metabolic synergy of breast cancer cells with tumor microenvironment: Interaction of breast cancer cells with normal fibroblasts, cancer-associated fibroblasts and adipocytes in vitro|
|Authors : Cheppail Ramachanndran,Ashley Juan,Daniel Moy,Karl-Werner Quirin,Enrique Escalon,Ziad Khatib,Steven J. Melnick|
SDS-PAGE, sodim dodecyl sulfate-polyacrylamide gel electrophoresis
|Keywords : ATP , adenosine triphosphate|
|Manuscript Code : Tumor microenvironment (TME) plays an important role on the progression and metastasis of tumors because of metabolic coupling and interaction between non-malignant cells in the TME and cancer cells. We have investigated the role of cancer-associated fibroblasts (CAF), normal fibroblasts (NF) and adipocytes on the growth of breast carcinoma cells in vitro. Additionally, the anti-glycolytic effects of supercritical CO2 extract of mango ginger (Curcuma amada Roxb) (CA) on both conventional and reverse Warburg effects were studied in the monocultures of CAF (CAF-05), NF (CCL-110), adipocytes (HPAd) and breast cancer (MCF-7) cells and co-cultures of breast cancer cells with CAFs, NF and HPAd. CA was highly cytotoxic to CAF-05 and MCF-7 cell lines compared to CCL-110 and HPAd cell lines. CA inhibited ATP synthesis in monocultures and co-cultures, though the rate of inhibition is higher in co-cultures than MCF-7 monoculture. Although non-malignant cell lines (CAF-05, CCL-110 and HPAd) have lower levels of lactate synthesis than MCF-7 cells, inhibitory effect of CA is more pronounced in co-cultures of MCF-7 with CAF-05, CCL110 and HPAd cells, respectively than MCF-7 cells alone. Gene expression studies showed that although transcripts of glycolysis-associated genes HIF-1α, MCT1, MCT4, Caveolin-1 and GLUT1 were present in non-malignant monocultures, protein translation of these genes were absent. Treatment of co-cultures of MCF-7+CAF-05 and MCF-7+HPAd cells with supercritical CO2 extract of mango ginger [Curcuma amada Roxb. (CA) - 5 g/ml] down regulated MCT1, MCT4, and Caveolin-1 proteins. CA treatment also inhibited HIF-1α and GLUT1 protein expression in co-cultures of MCF-7+HPAd cells. These results indicate that CA inhibits both Warburg and reverse Warburg effects in TME.|
|Title : Synergistic effect of supercritical CO2 extract of mango ginger (Curcuma amada Roxb.) with glycolytic inhibitors in human glioblastoma cells in vitro|
|Authors : Cheppail Ramachandran|
Glycolysis, Mango Ginger, ATP, Lactate, Gene Expression, Glioblastoma : Background: Cancer cells, unlike normal cells, principally use aerobic glycolysis with reduced mitochondrial oxidative phosphorylation for glucose metabolism, a phenomenon referred to as the Warburg effect. Glioblastoma, one of the most aggressive, lethal and incurable human tumors with a survival rate of 12-15 months in patients undergoing standard of care treatment involving surgery, chemotherapy and radiation therapy, has been shown to have a preferential metabolism of glucose through aerobic glycolysis. The cytotoxic effects of a previously described supercritical CO2 extract of mango ginger (Curcuma amada-CA) with two glycolytic inhibitors [2-deoxy-D-glucose (2-DG) and sodium oxamate (SO)] was investigated in the U-87MG human glioblastoma cell line. Methods: Cytotoxicity assay was performed with increasing concentrations of CA, 2-DG and SO as single agents and in combinations in U-87MG glioblastoma cells by the MTT assay. The cytotoxicity data was analyzed using CompuSyn software to determine the synergism/additive effect/antagonism between drugs. The effect of CA and glycolytic inhibitors on ATP and lactate synthesis was analyzed to establish the inhibitory effects of individual drugs as well as their combinations on glycolysis pathway. The modulatory effect of CA, 2-DG and SO as single agents or combinations on mRNA and protein expression of apoptotic and metastasis genes were also analyzed by RT-PCR and western blot hybridization, respectively. Results: The hexokinase inhibitor 2-DG and the lactate dehydrogenase-A inhibitor SO, both inhibiting the glycolysis pathway, showed synergistic cytotoxic effects with CA in the glioblastoma cell line with combination index values of <1 in the CompuSyn analysis. CA inhibits cellular ATP synthesis in a dose-dependent manner and it has better inhibition profile than 2-DG and SO. CA inhibits cellular lactate synthesis significantly better than 2-DG and SO at low concentrations, and CA+2-DG combination appears to be better than single agents at low doses for lactate inhibition in glioblastoma cells. Gene expression analysis by RT-PCR and western blot hybridization showed that CA, 2-DG and SO as well as their combinations up regulate the ratio of Bax/Bcl-2, p21, TIMP1 and caspase-3 expression and down regulate mutant p53 and MMP2 expression that may increase apoptosis and inhibit cell proliferation as well as metastasis of tumor cells. Conclusion: The combination of CA with glycolytic inhibitors like 2-DG and SO is beneficial for inhibition of growth, proliferation and migration of glioblastoma cells. These in vitro results support the rationale for conducting in vivo studies combining CA with 2-DG and SO in human glioblastomas.
|Keywords : Glycolysis , Mango Ginger , ATP , Lactate , Gene Expression , Glioblastoma : Background: Cancer cells , unlike normal cells , principally use aerobic glycolysis with reduced mitochondrial oxidative phosphorylation for glucose metabolism , a phenomenon referred to as the Warburg effect. Glioblastoma , one of the most aggressive , lethal and incurable human tumors with a survival rate of 12-15 months in patients undergoing standard of care treatment involving surgery , chemotherapy and radiation therapy , has been shown to have a preferential metabolism of glucose through aerobic glycolysis. The cytotoxic effects of a previously described supercritical CO2 extract of mango ginger (Curcuma amada-CA) with two glycolytic inhibitors [2-deoxy-D-glucose (2-DG) and sodium oxamate (SO)] was investigated in the U-87MG human glioblastoma cell line. Methods: Cytotoxicity assay was performed with increasing concentrations of CA , 2-DG and SO as single agents and in combinations in U-87MG glioblastoma cells by the MTT assay. The cytotoxicity data was analyzed using CompuSyn software to determine the synergism/additive effect/antagonism between drugs. The effect of CA and glycolytic inhibitors on ATP and lactate synthesis was analyzed to establish the inhibitory effects of individual drugs as well as their combinations on glycolysis pathway. The modulatory effect of CA , 2-DG and SO as single agents or combinations on mRNA and protein expression of apoptotic and metastasis genes were also analyzed by RT-PCR and western blot hybridization , respectively. Results: The hexokinase inhibitor 2-DG and the lactate dehydrogenase-A inhibitor SO , both inhibiting the glycolysis pathway , showed synergistic cytotoxic effects with CA in the glioblastoma cell line with combination index values of <1 in the CompuSyn analysis. CA inhibits cellular ATP synthesis in a dose-dependent manner and it has better inhibition profile than 2-DG and SO. CA inhibits cellular lactate synthesis significantly better than 2-DG and SO at low concentrations , and CA+2-DG combination appears to be better than single agents at low doses for lactate inhibition in glioblastoma cells. Gene expression analysis by RT-PCR and western blot hybridization showed that CA , 2-DG and SO as well as their combinations up regulate the ratio of Bax/Bcl-2 , p21 , TIMP1 and caspase-3 expression and down regulate mutant p53 and MMP2 expression that may increase apoptosis and inhibit cell proliferation as well as metastasis of tumor cells. Conclusion: The combination of CA with glycolytic inhibitors like 2-DG and SO is beneficial for inhibition of growth , proliferation and migration of glioblastoma cells. These in vitro results support the rationale for conducting in vivo studies combining CA with 2-DG and SO in human glioblastomas.|
|Title : Pattern of delayed presentation of breast cancer patients: Evidence from Rangpur Medical Hospital, Rangpur, Bangladesh|
|Authors : Bimal Chandra Roy,Saimun Naher,Md. Shafiul Alam,Md. Abu Hanifa,Prosannajid Sarkar|
Background: Bangladesh is facing a high burden of breast cancer disease. It is the 2nd leading cause of cancer death after cervical cancer. Delay in seeking treatment contributes in breast cancer patients’ presentation at the tertiary hospitals when cancer is at an advanced stage and leads to poor prognosis. Late presentation with advanced stage is the common feature of breast cancer patients of Bangladesh. Objectives: The main objective of this study is to explore factors associated with delay in seeking cancer treatment among breast cancer patients at a tertiary care Hospital. Methods: Cross sectional observational study was conducted in the Department of Surgery Rangpur Medical College Hospital, Rangpur from January 2013 to December 2014. Patients who admitted to surgery ward and fulfill inclusion criteria during my study period was included in this study. Sixty two patients with carcinoma breast were included in this study. Thorough search for personal information and reasons behind delayed presentation was made from the patients. Information regarding tumor stage at time of diagnosis was extracted from hospital records and involved pathologic and clinical data. The data was collected in a pre designed data collection sheet. Correlation between personal and social factors and delayed presentation were made to find out the factors responsible for delayed presentation. Results: 36% patients came within 3 months of noticing the lump in breast and 64% delayed more than 3 months. Among 62 patients 76% were from low socioeconomic class, 62% had no formal primary education and 74% took initial treatment from Homeopath and traditional healer. Significant association was found with these patient related factors and delayed presentation. Association was also found between delayed presentation and advanced stage of disease. Conclusion: The findings suggest that 64% women with breast cancer presented late and have significant effect on their disease prognosis. There is a great impact of patient factors (personal and social) on delayed presentation. To reduce patient delay community awareness, health education programs regarding breast cancer should be implemented for target women who are at higher risk of delay.
|Keywords : Breast cancer , Pattern , delayed.|
|Title : Carbon Nanotubes- the Holy Grail in Anticancer Therapy|
|Authors : Varsha Tripathi,Manoj Kumar Mishra,Pallavi Tiwari,Mayank Srivastava,Saurav Ghoshal|
Carbon nanotubes(CNT) are allotropes of carbon with cylindrical nanostructures. They can be visualized as a sheet of carbon atoms rolled up into a tube with a diameter of around tens of nanometers. They have electrical, optical, thermal, mechanical properties. There are two types of CNTs, single walled CNTs and multi walled CNTs. The multiwalled CNTs are formed by several concentric layers of rolled graphite. It has good adsorption properties which can detect some chemicals and biological agents. It is used to transport drug as well as proteins, DNA, RNA, into cells. The carbon network of shell is considered to be a result of the arrangement of carbon atoms in graphite sheets. Some properties of CNTs such as ease of cellular uptake, high drug loading, thermal ablation, among others, render them useful for cancer therapy. Cancer is one of the most challenging diseases of modern times because its therapy involves distinguishing normal healthy cells from affected cells. CNTs may prove to be the Holy Grail in cancer therapy because phenomena such as EPR, allow CNTs to distinguish normal cells from affected ones,. Considerable work has been done on CNTs as drug delivery systems over the last two decades. However, concerns over certain issues such as biocompatibility and toxicity have been raised and warrant extensive research in this field.
|Keywords : Carbon Nanotubes , Anticancer , SWCNT , MWCNT , Drug Delivery|
|Title : Toxicological effect of plant flavonoids on MCF-7, A-549 and PA-1 cancer cell lines|
|Authors : Rajeev Nema,Sarita Khare|
The research aimed in the direction of five different medicinal plants (hydro alcoholics extract) to evaluate anticancer activity and cytotoxcity. The most important purpose of this study is to evaluate cytotoxicity of these medicinal plants with facilitate of MTT assay. Concentrations are prepared of each plant extracts which are 100 μg/ml, 10 μg/ml, 0.1 μg/ml, 0.01 μg/ml and 5-10×103 cells/ml are taken into each well which are exposed to different Concentrations of plant extracts crude and elute of column chromatography [CC] for 96 hrs and then treated with MTT and take absorbance at 570 nm. Standard drug was used for IC50 of Doxorubicin MCF-7 500nm, A549- 550nm, PA-1- 580nm.
|Keywords : Toxicology , Medicinal Plants|
|Title : Li-Fraumeni syndrome in a patient with familial hyperlipidemia from Western Iran, a case report|
|Authors : Mehrdad Payandeh,Reza Khodarahmi, Masoud Sadeghi,Edris Sadeghi|
Mutations of germ-cell line TP53 gene are mainly reported in Li–Fraumeni syndrome(LFS). LFS-associated breast cancers are both hormone receptor and human epidermal growth factor receptor 2 positive. The purpose of this study is presentation of one case of LFS, associated with familial hyperlipidemias. A 52-year-old woman referred to clinic of oncology with a pain in the left breast. Her pathology report showed that she had breast cancer and computed tomography scan showed no evidence of metastasis. Markers of estrogen receptor, progesterone receptor and p53 were positive, IHC3+ and Ki67 in 20% of cells. We report the first case of a LFS patient with breast cancer and familial hyperlipidemias in Iran who ER, PR, P53 and HER-2 markers for her are positive. Also it is probably that LFS-associated cancer and atherosclerosis diseases are related to each other.
|Keywords : Germ-Line Mutation , Hyperlipidemias , Li-Fraumeni Syndrome|
|Title : Cancer –From the Horizons of Unani Medicine|
|Authors : Amena Khatoon,Mohd azeem|
Cancer is an abnormal growth of cells which tend to proliferate in an uncontrolled way and in some cases to metastasize.cancer is the Latin word for crab .The ancients used the word to mean malignancy because of the crab like tenacity.It is a mass of tissue formed as a result of abnormal, excessive, uncoordinated, autonomous and purposeless proliferation of cell. Hippocrates [460-377BC] coined the term karkinos for cancer of the breast.. Cancer cells in other words refuse to stop multiplying and continue to increase in number. It is the failure to stop multiplying which is the hallmark of a cancer. This they do even at the cost of other normal cells of the body which are starved to death for lack of nutrition. In Unani literature the basic cause of disease is change in akhlat with respect to their quality and quantity. When the black bile (sauda) deviates from its normal quantity and quality.
|Keywords : Akhlat ,Sauda ,Unani Literature|
|Title : Association between ABO Rh Blood Groups and Oral Potentially Malignant Disorders|
|Authors : Jyoti R Byakodi,K Pushpangali|
Background: A number of potentially malignant disorders like leukoplakia, erythroplakia, oral submucous fibrosis constitute a detectable pre-clinical phase of oral cancer. There is a high prevalence of these lesions in India, which is attributed to culture, ethnic, geographic factors, low socio-economic status and varying lifestyle factors. The genetic relation of these lesions is less studied. Blood groups play an important role in the immunogenetic system. Very few studies have been done in India to see the association between blood group and potentially malignant disorders. Hence the present study was under taken to assess the relationship between ABO Rh blood group and oral potentially malignant disorders. Materials and methods: 154 cases with oral potentially malignant disorders diagnosed based on the WHO criteria (1980) were included in the study. Patients reporting to M.S.Ramaiah Dental and Hospital, Bangalore and Kaiwara center (a peripheral outreach center) were included in the study after obtaining an informed consent. Information regarding socio-demographic details was collected using proforma and blood group was assessed using blood group kit with Rapid slide test method. Statistical analysis was done using SPSS software11. Descriptive statistics and chi square test were used to find the correlation. Result: Out of 154 cases, 88(57.14%) were males and 66(42.86%) females. 84(54.5%) cases were leukoplakia, 2(1.3%) cases were erythroplakia, 53(34.4%) cases were OSMF and 15(9.7%) cases were with multiple oral potentially malignant disorders. 57(37.01%) of cases belonged to B+ve blood group followed by 37(20.43%) belonged to A+ve blood group.
|Keywords : Oral Potentially Malignant Disorders , Leukoplakia , Erythroplakia , Oral Submucous Fibrosis , Blood Group|
|Title : Analysis of gene expression in normal and oncogene transfected cells of rat Model|
|Authors : G. Renuga,J. Amba Bhavani|
Cells in culture offer many opportunities to study the characteristics of both the normal and cancer cells. In order to understand the molecular mechanisms of cell proliferation, an attempt has been made to study the proteins which are at different phases of the cell cycle. To achieve this objective, rat embryo fibroblasts were synchronized at Go phase by serum starvation for 72h and then stimulated with serum mitogens or purified growth factors. The newly synthesized proteins were labeled with [ 35 S] methionine at different phases of the cell cycle after stimulation and the secreted proteins were analyzed by SDS- polyacrylamide gels. One of the proteins which has shown to be involved in growth regulation was purified and bioassays were carried out to determine its function. The protein 48 KDa was found to be a major compound of the extracellular matrix (ECM) whereas the protein 26 KDa was not a matrix associated protein. When the cells were arrested at G1/S boundary with hydroxyurea (HOU), high levels of protein with 45KDa protein was observed in the medium. Normal rat embro fibroblasts were transfected with myc and ras oncogenes and the transformed colonies were cultured and purified 45KDa protein inhibited the DNA synthesis of myc and ras oncogene transformed cells in which the 45KDa protein secretion was down regulated, indicating the inhibition of DNA synthesis.
|Keywords : Cell cycle , DNA synthesis , growth factor , Protein , Oncogene , rat embryo fibroblasts|
|Title : Histopathological study of tumours of cervix|
|Authors : Atul Jain|
Carcinoma of cervix is the most common cancer in Indian women and accounts for 20% of all malignant tumours in the females. Squamous cell carcinomas (SCC) account for 75-80%, adenocarcinoma 15-25%, and adenosquamous carcinomas 3-5% of cervical cancers. Objectives: To study histopathology of carcinoma of cervix and to find out the incidence rate, most common type of the carcinoma of cervix and to find out age predilection in various types of carcinoma of cervix. Material and Method: This study was undertaken in the department of pathology over a period of 2 years from May 2010 to April 2012. All Hysterectomy specimens and cervical biopsies were processed routinely and paraffin sections were taken and stained with Haematoxylin and Eosin (H&E) for microscopic examination. Tumours of the cervix were classified and studied according to WHO classification. Results: The most frequent condition diagnosed on cervical biopsies and hysterectomy specimens were benign cervical polyp (66.3%), followed by carcinoma (23.7%) and squamous intraepithelial lesion (0.2%). Out of all benign cervical polyps, occurrence of adenomatous polyp was highest. Incidence of squamous cell carcinoma (19.6%) was more as compared to adenocarcinoma (3.7%). The mean age of the squamous cell carcinoma, adenocarcinoma, squamous intraepithelial lesion and benign cervical polyp was 49.1 years, 43.5 years, 47.7 years and 44.6 years respectively. Conclusion: Incidence of squamous cell carcinoma was more as compared to adenocarcinoma. Occurrence of malignant lesions was earlier during 4th decade as compared to benign cervical lesions.
|Keywords : Cervical polyp , Carcinoma cervix , Squamous cell carcinoma , Adenocarcinoma|
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- International Committee of Medical Journal Editors. Uniform Requirements for Manuscripts Submitted to Biomedical Journals: Writing and Editing for Biomedical Publication. 2006.
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