Search

Natural killer (NK) cells are one of the major components of innate immunity, with the ability to mediate antitumor activity. Understanding the role of NK-cell–mediated tumor killing in controlling of solid tumor growth is still in the developmental stage. We have shown recently that bitter melon extract (BME) modulates the regulatory T cell (Treg) population in head and neck squamous cell carcinoma (HNSCC). However, the role of BME in NK-cell modulation against HNSCC remains unknown. In this study, we investigated whether BME can enhance the NK-cell killing activity against HNSCC cells. Our results indicated that treatment of human NK-cell line (NK3.3) with BME enhances ability to kill HNSCC cells. BME increases granzyme B accumulation and translocation/accumulation of CD107a/LAMP1 in NK3.3 cells exposed to BME. Furthermore, an increase in cell surface expression of CD16 and NKp30 in BME-treated NK3.3 cells was observed when cocultured with HNSCC cells. Collectively, our results demonstrated for the first time that BME augments NK-cell–mediated HNSCC killing activity, implicating an immunomodulatory role of BME. Cancer Prev Res; 10(6); 337–44. ©2017 AACR.

Background: Screening using fecal occult blood testing (FOBt) reduces colorectal cancer mortality, but the test has low sensitivity. A “missed” cancer may cause psychologic harms in the screened population that partially counteract the benefits of early detection. Methods: Three hundred and eleven people diagnosed with colorectal cancer (i) after a negative FOBt result (interval cancer), (ii) a positive result (screen-detected cancer), or (iii) in regions where screening was not offered, completed questions on quality of life (FACT-C), depression (CES-D), perceived diagnostic delay, and trust in the results of FOBt screening. Fifteen withheld consent to data matching with medical records, leaving a sample size of 296. Results: Controlling for demographic and clinical variables, patients with an interval cancer reported poorer quality of life (difference in means = 6.16, P = 0.03) and more diagnostic delay (OR, 0.37; P = 0.02) than patients with screen-detected disease, with no differences in depression. No differences were observed between the interval cancer group and the group not offered screening on these measures. Patients with an interval cancer reported the lowest levels of trust in FOBt. Conclusions: An interval cancer has adverse effects on trust in FOBt, but does not result in worse psychologic outcomes compared with people diagnosed in areas with no screening program. People with an interval cancer report poorer quality of life than people with screen-detected disease. Impact: Improvements in test sensitivity could improve quality of life among people who complete an FOB test over and above any benefits already conferred by earlier detection. Cancer Epidemiol Biomarkers Prev; 24(7); 1–7. ©2015 AACR .

Head and neck squamous cell carcinoma (HNSCC) represents one of the most common malignancies worldwide with a high mortality rate mainly due to lack of early detection markers, frequent metastasis and aggressive phenotype. Recently, long non-coding RNAs (lncRNAs) have been shown to have important regulatory roles in human cancers. Up-regulation of prostate cancer-associated transcript 1 (PCAT1) in TCGA dataset of HNSCC was noted. In this study, we aimed to evaluate role of the PCAT1 in HNSCC. During validation, we observed up-regulation of PCAT1 in HNSCC patient samples and cell lines. siRNA mediated knock-down of PCAT1 significantly reduced cell proliferation, sphere formation and wound healing in HNSCC cell lines (JHU29 and Cal27). Mechanistic study revealed a significant down-regulation of c-Myc and AKT1 in both RNA and protein levels upon knockdown of PCAT1 in HNSCC cell lines. In-silico analysis suggested a significant positive correlation between PCAT1 and c-Myc or PCAT1 and AKT1 in HNSCC cell lines. Subsequently, we observed modulation of MAPK signalling by activation of p38 and apoptosis signal-regulating kinase 1 (ASK1) upon knockdown of PCAT1, and induction of apoptosis as evident of caspase 9 activation and PARP cleavage. Introduction of PCAT1 into HNSCC xenograft tumors displayed a reduction in tumor volume. Together our data demonstrated an important role of the PCAT1 in HNSCC progression and has potential as a therapeutic candidate. Citation Format: Subhayan Sur, Robert Steele, Ratna B. Ray. Depletion of PCAT1 in head and neck cancer cells limits tumor growth by regulating c-Myc, AKT1 and MAPK signalling pathways [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1813.

Prostate cancer remains the second leading cause of cancer deaths among American men. Early diagnosis increases survival rate in patients; however, treatments for advanced disease are limited to hormone ablation techniques and palliative care. Thus, new methods of treatment are necessary for inhibiting prostate cancer disease progression. Here, we have shown that miRNA-29b (miR-29b) expression was lower in prostate cancer cells (PC3 and LNCaP) as compared with immortalized prostate epithelial cells. Between these two prostate cancer cell lines, metastatic prostate cancer PC3 cells displayed lower expression of miR-29b. We also observed a significant downregulation of miR-29b expression in human prostate cancer tissues as compared with patient-matched nontumor tissues. PC3 cells ectopically expressing miR-29b inhibited wound healing, invasiveness, and failed to colonize in the lungs and liver of severe combined immunodeficient mice after intravenous injection, while PC3 cells expressing a control miRNA displayed metastasis. Epithelial cell marker E-cadherin expression was enhanced miR-29b transfected in prostate cancer cells as compared with cells expressing control miRNA. On the other hand, N-cadherin, Twist, and Snail expression was downregulated in PC3 cells expressing miR-29b. Together these results suggested that miR-29b acts as an antimetastatic miRNA for prostate cancer cells at multiple steps in a metastatic cascade. Therefore, miR-29b could be a potentially new attractive target for therapeutic intervention in prostate cancer. Mol Cancer Ther; 11(5); 1166–73. ©2012 AACR.

Breast cancer is the most common malignant cancer diagnosed in women worldwide each year. Triple-negative breast cancers (TNBC), defined by the lack of expression of estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2), account for approximately 15% of breast cancers. Moreover, TNBC is usually more aggressive and metastatic with a worse prognosis. Bitter melon (Momordica charantia) is well known as a dietary supplement. Recently, we have shown an anti-cancer activity of bitter melon extract (BME) in breast cancer mouse models, although the mechanism is poorly understood. Furthermore, we became interested to understand the role of BME on cholesterol and lipid metabolism in TNBC. Our mass spectrometry data suggested that stored as a cholesteryl esters (CE) in Lipid droplets (LD) were much lower in BME treated TNBC cells. We found that not only CE but also LD was reduced in BME treated TNBC cells. The acyl-CoA:cholesterol acyl transferase-1 (ACAT-1) catalyzes the formation of CE from free cholesterols together with long-chain fatty acyl coenzyme A. Therefore, inhibition of ACAT-1 resulted in downregulation of CE, and upregulation free cholesterols. The attenuated CE is known to reduce cancer proliferation. Subsequent study demonstrated that BME treatment highly inhibited ACAT-1 in TNBC cells. The upregulated free cholesterol negatively regulates sterol regulatory element-binding proteins (SREBPs), which regulates lipogenesis protein such as fatty acid synthase (FASN) associated with cancer cell proliferation. We also observed that SREBP-1 and -2 were barely detectable, and FASN and low-density lipoprotein receptor (LDLR) were reduced in BME treated TNBC cells as compared to control cells. To our knowledge, this is the first report demonstrating that BME suppresses TNBC cell growth through ACAT-1 inhibition. BME supplement may potentially be an additional regimen in human breast cancer treatment. Citation Format: So Hee Shim, Robert Steele, Ratna Ray. Bitter melon extract suppresses triple-negative breast cancer growth by inhibiting ACAT1 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3532.

Oral squamous cell carcinoma (OSCC) is a complex malignancy and tobacco habit is one of the main etiological factors. Our previous study showed potential chemopreventive effect of bitter melon (Momordica charantia) extract (BME) in different pre-clinical cancer models like head and neck, breast, prostate. The aim of this study is to understand the molecular mechanism of BME in prevention of carcinogen induced in-vivo oral carcinogenesis. For this purpose, we induced oral cancer in immunocompetent mice by using tobacco associated carcinogen 4-nitroquinoline 1-oxide (4-NQO), and treated with BME through their drinking water. We observed that BME feeding significantly reduced the incidence of 4-NQO induced oral cancer. Histological analysis showed moderate dysplasia to invasive squamous cell carcinoma in 4NQO treated mice tongue, whereas no remarkable changes were seen in the BME-fed mice. RNA-seq data showed differential expression of several genes during carcinogenesis and during prevention of the cancer. We previously showed that BME treatment decreases the infiltrating regulatory T (Treg) cells by inhibiting FoxP3+ populations in the tumors and in spleens, suggesting an immunomodulatory role of BME in prevention. Here, we observed elevated expression of pro-inflammatory genes s100a9, IL23a, IL-1β and immune check point gene PDCD1/PD1 of the pathway, during oral cancer development. Interestingly, BME treatment significantly reduced the expression of these molecules. Subsequently, we observed the enhancement of MMP9 during carcinogenesis, which was reduced in BME-fed mouse tongue tissues. Taken together, our observation strongly demonstrated a chemopreventive effect of BME on carcinogen induced oral cancer. Citation Format: Subhayan Sur, Robert Steele, Ratna B. Ray. Bitter melon extract modulates immune signaling in prevention of tobacco associated carcinogen induced oral carcinogenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5203.

Lung cancer is the leading cause of cancer death among both men and women. Only ∼15% of people diagnosed with non–small cell lung cancer (NSCLC) survive this disease beyond 5 years. Thus, novel therapeutic strategies are urgently needed to improve the clinical management of this devastating disease. We have previously shown the antiproliferative effect of MBP-1 on several human cancer cells. In this study, we have examined the potential of MBP-1 as a gene therapeutic candidate in regression of non–small cell lung tumor growth. We have observed that exogenous expression of MBP-1 in NSCLC cells (H1299) induces massive cell death. To determine the gene therapeutic potential of MBP-1, replication-deficient recombinant adenovirus expressing MBP-1 was given intratumorally in human lung cancer xenografts in nude mice. Our results showed a significant regression of lung tumor growth and prolonged survival on treatment with MBP-1 compared with the control groups (saline or dl312). Subsequently, the mechanism of MBP-1–mediated H1299 cell death was investigated. Our results suggested that MBP-1 induced poly(ADP-ribose) polymerase cleavage in H1299 cells; however, treatment with pan-caspase inhibitor did not protect against MBP-1–induced cell death. Cells transduced with MBP-1 displayed early plasma membrane permeability, mitochondrial damage without cytochrome c release, and extensive cytoplasmic vacuolation, yielding a morphotype that is typical of necrosis. Taken together, this study suggests that MBP-1 expression induces a novel form of necrosis-like cell death and MBP-1 could be a potential gene therapeutic candidate against non–small cell lung tumor growth. (Cancer Res 2006; 66(24): 11907-12)

Prostate cancer is the most frequently diagnosed cancer in men and the second leading cause of male cancer death in the United States. Early detection and improved procedures for surgical intervention and radiation therapy have reduced the fatalities; however, there is no effective cure for men with advanced disease and additional therapy is urgently needed. We have previously shown that MBP-1 acts as a general transcriptional repressor and exerts an antiproliferative effect on several human cancer cells. MBP-1 possesses two repressor domains, located at the amino and carboxyl termini. In this study, we have examined the potential of the repressor domains of MBP-1 as a gene therapeutic candidate in regression of prostate tumor growth. Our results suggested that replication-deficient adenovirus-mediated delivery of amino-terminal (MBP-AR) or carboxyl-terminal (MBP-CR) repressor domain of MBP-1 exerted an antiproliferative effect, like the full-length MBP-1, and induced caspase-independent apoptosis in prostate cancer cells. Next, we investigated the therapeutic effectiveness of MBP-1 repressor domain on prostate tumors. When tested in human tumor xenografts in nude mice, MBP-CR suppressed prostate tumor growth more effectively than full-length MBP-1, whereas MBP-AR delayed prostate tumor growth. Together, these results suggested that MBP-CR expression has an antiproliferative effect in human prostate cancer cells, being more effective than the full-length MBP-1 in preventing tumor growth.

Purposes of the study: Head and neck squamous cell carcinoma (HNSCC) is the sixth most widespread cancer and leading cause of cancer related death world widely. Regardless of the advancement in treatment processes the overall survival rate of patients having this cancer has not substantially improved in the past few decades. Development of tumor and its progression exclusively resides in a small population of cancer cells, termed as cancer stem cells (CSCs). This population of cells can self-renew and re-form the ordered organization of tumors. CSCs contribute to HNSCC and may offer prominent therapeutic strategy for the management of HNSCC. We previously observed that c-Fos was highly up-regulated in the HNSCC sphere forming cells. In the present study, we explored the role of c-Fos in the tumor initiation in relation with cancer stemness and EMT. Experimental procedures: We examined whether overexpression of c-Fos has an effect on tumorigenicity and stemness. For this, c-Fos was overexpressed in non-tumorigenic HNSCC cell lines and tumor growth was investigated. We also examined the expression of different CSC marker genes expression by Western blot analysis. Summary: Our results demonstrated that exogenous expression of c-Fos in non-tumorigenic MDA1386Tu cells makes the cells tumorigenic in nude mice. Mechanistic investigations showed that c-Fos overexpression in MDA1386Tu cells enhanced epithelial-mesenchymal transition (EMT)/CSC properties and increased numbers of oralspheres when grown in ultra-low binding plate. Conclusions: Collectively, our data strongly demonstrate that c-Fos plays a critical role in the modulation of EMT and cancer stem cell reprogramming in HNSCC cells and enhance tumor growth. Note: This abstract was not presented at the meeting. Citation Format: Naoshad Muhammad, Sourav Bhattacharya, Robert Steele, Nancy Phillips, Ratna B. Ray. C-Fos drives non-tumorigenic head and neck squamous cell carcinoma cells to tumorigenic by promoting cancer stem cells like properties [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5761. doi:10.1158/1538-7445.AM2017-5761

The purpose of the study: Hepatocellular carcinoma (HCC) is global problem and incidence of HCC is increasing in last several years. Hepatitis C virus (HCV) infection is one of the major cause towards development of HCC. HCV infection is highly prevalent in African American population compared to other ethnic groups and these people are less likely to naturally clear HCV. Higher incidence of HCC and mortality was noted in HCV infected African Americans as compared to Caucasians. However, the explanation for this disparity and the molecular mechanisms behind this are currently unknown. Circulating microRNAs in the blood are emerging as biomarkers for pathological conditions. Differential expression of miRNAs among ethnic groups would be important for optimizing personalized treatment strategies. Experimental procedures: In this study, we assessed the differential expression of circulatory miRNAs from HCV infected African Americans and Caucasians by cancer specific miRNA array profiling. Expression of significantly altered miRNAs was validated by qRT-PCR. Results: We identified increased expression of miR-146a, miR-150 and miR-155 in HCV infected African Americans patient sera as compared to sera of Caucasians. Further analysis demonstrated that these miRNAs were significantly elevated in African Americans diagnosed with HCV-mediated HCC. Higher expression of miR-150 was also noted in cirrhosis and HCC in African Americans, which may serve as predictor of liver disease progression in this population. Conclusion: The differential expression of miRNAs suggests that these miRNAs and their target genes could be useful to gain further mechanistic insight of racial disparity associated with HCV pathogenesis. Citation Format: Pradip Devhare, Robert Steele, Adrian M. Di Bisceglie, David E. Kaplan, Ratna B. Ray. Circulatory microRNA expression profiling between HCV-infected African & Caucasian Americans: implications for racial health disparities [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5724. doi:10.1158/1538-7445.AM2017-5724

Hepatocellular carcinoma (HCC) is the world's third most common cancer and causes 600,000 deaths annually. Early detection is the key for improving the survival of HCC patients. Current diagnostic markers, such as alpha fetoprotein or fibroscan have their own limitations. Investigating serum microRNA as biomarkers for early diagnosis of cancer has become an important tool in clinical research. In this study, we tested the hypothesis that serum microRNAs (miRNAs) can be used as a potential biomarker for HCC. We analyzed serum miRNA using reverse transcriptase-quantitative PCR (qRT-PCR) from serum samples. Our study revealed that two miRNAs, miR-30e and miR-223, were expressed at significantly lower levels (P < 0.003) in the sera of HCC patients as compared to healthy volunteers. Further, expression of these miRNA was compared between sera from chronic liver disease (CLD) and HCC patients. miR-30e and miR-223 expression was significantly lower in HCC sera as compared to sera from CLD patients. Both miRNA expression levels were lower in HCC liver biopsy specimens as compared to normal liver RNA. Taken together, our results indicated that serum miR-30e and miR-223 are useful biomarkers of HCC irrespective of etiology. Citation Format: Ratna B. Ray, Sourav Bhattacharya, Robert Steele, Adrian M. Di Bisceglie, Sounak chakraborty. Serum miR-30e and miR-223 are promising non-invasive biomarkers for hepatocellular carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1085.

Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer and leading cause of cancer related mortality worldwide. Despite the advancement in treatment procedures the overall survival rate of patients has not considerably enhanced in the past few decades. Therefore, new strategies to achieve a favorable response for the improvement in the prognosis of HNSCC are urgently needed. In this study we examined the immune modulatory activity of bitter melon extract (BME) in HNSCC tumor microenvironment. For this, mouse head and neck cancer (SCCVII) cells were subcutaneously injected into the flanks of C3H mice. BME was fed orally, and tumor growth was evaluated. We observed that BME treatment significantly inhibits the tumor growth in BME fed mice as compared to control group. Flow cytometry data showed that BME treatment not only decreases the infiltrating T regulatory (Treg) cells in the tumor but also in splenocytes. Additionally, BME treatment reduces Th17 cell population in the tumor and augments in the spleen. However, BME treatment did not alter Th1 and Th2 populations. Further study suggested that BME inhibits cell proliferation as evident from low expression of PCNA and c-Myc in the tumors of BME fed mice as compared to that of control group. To our knowledge, this is the first report showing BME exerts immunomodulatory effect in regressing HNSCC tumor growth in preclinical model. Citation Format: Sourav Bhattacharya, Robert Steele, Naoshad Muhammad, Guangyong Peng, Ratna B. Ray. Immunomodulatory activity of bitter melon extract in head and neck squamous cell carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5125.

Prostate cancer remains the second leading cause of cancer deaths among American men. Earlier diagnosis increases survival rate in patients. However, treatments for advanced disease are limited to hormone ablation techniques and palliative care. Thus, new methods of treatment and prevention are necessary for inhibiting progression of the disease to a hormone refractory state. For centuries, Ayurveda has recommended the use of bitter melon (Momordica charantia) as a functional food to prevent and treat human health related issues. We have recently discovered an anti-proliferative effect of bitter melon extract (BME) in a number of cancer cell lines, including prostate, without affecting normal cell growth. Treatment of prostate cancer cells with bitter melon extract modulated Bax expression and induced poly(ADP-ribose) polymerase cleavage. TRAMP (transgenic adenocarcinoma of mouse prostate) male mice display prostatic intraepithelial neoplasia (PIN) at 10-12 weeks of age. We examined whether oral administration of BME as a dietary compound inhibits or delay the conversion of normal prostate to PIN in these mice. We have observed prevention of PIN in BME-fed mice as compared to untreated control. Further tissues analyses for mechanism of cancer prevention are in progress and the results will be discussed. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4618. doi:10.1158/1538-7445.AM2011-4618