International Journal of Cancer Research and Molecular Mechanisms

3

Authors: J Younus

Background: Explaining targeted therapies to oncology patients is a challenging task for most health care professionals (HCPs). We opted to explore a unique pictorial approach with Apps for patient education. Design/Methods: Three applications (Apps), one each for Tamoxifen, Aromatase inhibitors (AI’s) and Trastuzumab were created, utilizing the iPhone/iPad platform. The content of these Apps was approved through Breast Disease Site Team and Patient Education Committee. The study was approved by the Ethics Committee at the Western University. After obtaining consent, the Apps with animated cartoons were verbally reviewed with adult patients with breast cancer by the HCP. A patient satisfaction survey and two “exam” questions with a multiple-choice answer format were completed after each App was used by a HCP. Results: A total of 64 patients participated, 33 with the AI App, 19 with the tamoxifen App and 12 with the trastuzumab App. For the “exam questions” segment, there were no wrong answers given. The vast majority of patients rated the Apps very highly on all questions with the mean for these questions ranging between 6.67 to 7 on the visual analogue scale. Conclusions: The use of Apps is a novel and effective approach to educate oncology patients regarding molecularly targeted treatments. These Apps are quick, easy to use, with smart phones or an I-pad. From our results, it appears that patients liked this approach and actually understood the contents explained to them.

6

Authors: Okumura T

Background: Tricho-rhino-phalangeal syndrome-1 (TRPS1), a multi zinc-finger nuclear protein, has been reported to regulate cell proliferation, epithelial-to-mesenchymal transition (EMT), differentiation and apoptosis during development and tumor progression. The aim of this study was to investigate clinico-pathological significance of TRPS1 expression in gastric cancer (GC). Materials and Methods: The expression of TRPS1 was detected by immunohistochemistry in 110 surgically resected GC specimens using a tissue microarray (TMA). The patients consisted of 78 males and 32 females. Average age was 68.4 years old. The TNM stages of the patients were as follows: stage I, 17 patients; stage II, 34; stage III, 36; and stage IV, 23. After careful examination by two independent researchers, the immunohistochemical results for these patients were scored according to intensity and distribution. We additionally obtained surgically removed specimens from 10 gastric cancer patients to immunohistochemically detect the expression of TRPS-1, as well as to extracted RNA to detect the expression of microRNA (miR) such as miR221 and miR222. Results: Among the 110 patients with GC, 30 patients (27.3%) and 80 patients (72.7%) showed high and low expression of TRPS1, respectively. Low expression of TRPS1 was associated with distant metastasis (p=0.01). Age, gender, depth of the tumor, lymph node metastasis, TNM stage and histological grade were not associated with TRPS1 expression. The overall and cause-specific survivals of the TRPS1-low patients were significantly worse than those of the TRPS1-high patients (Wilcoxon p=0.043 and p=0.048, respectively). However, TRPS1 expression was not an independent prognostic factor of the patients. In the 10 surgically resected gastric cancer tissues, the lower expression of TRPS1 was significantly associated with higher expression of miR221 (p=0.017) and miR222 (p=0.007). Conclusion: Down regulation of TRPS1 expression was associated with distant metastasis and was an unfavorable prognostic factor in patients with GC, providing us with a novel prognostic marker and a potential therapeutic target. miR221 and miR222 were suggested to have a role in regulation of TRPS1 expression in GC.

11

Authors: Marsha Crochiere

Non-small cell lung cancer (NSCLC) has poor prognosis even with various treatment options. Unfortunately, resistance develops quickly and novel therapies are needed. Exportin-1 (XPO1) is a nuclear export protein upregulated in many types of cancers. Inhibition of XPO1 by selinexor (KPT-330), an oral Selective Inhibitor of Nuclear Export (SINE) compound, leads to nuclear accumulation of tumor suppressor proteins (TSPs), cell cycle arrest and cancer cell death. Selinexor is being evaluated in Phase I and II clinical trials in many different cancer indications (see clinicaltrials.gov for details) including lung cancer (NCT02351505). To date, selinexor has been given to >1400 patients and has shown good tolerability with manageable side effects. In this study the effects of selinexor on NSCLC cell growth and apoptosis were evaluated in vitro and in vivo. Selinexor inhibited tumor cell growth and clonogenic formation and induced cell cycle arrest and apoptosis in cell lines regardless of genetic background (EC50: 25-700 nM). The XPO1 cargo proteins p53, p21, IκB, E2F4, and survivin demonstrated strong nuclear localization after 4 to 24 hour treatment with selinexor followed by apoptosis (i.e. PARP, caspases-3 and -8 cleavage). Selinexor altered the localization of the NF-κB inhibitor IκB and functional evaluation of NF-κB in A549 and NCI-H1299 revealed transcriptional repression with EC50 values similar to the inhibition measured in cell proliferation assays. A549 xenografts in mice treated with selinexor showed markedly greater tumor growth inhibition (%TGI=81% at 20 mg/kg selinexor vs vehicle, p<0.0001) versus cisplatin-treated mice (%TGI=13% at 5 mg/kg cisplatin vs vehicle, p=0.06) when compared to vehicle. Using IHC on paraffin embedded tissue, treated tumors showed reduced cell proliferation (Ki67) while inducing nuclear accumulation of p53, p21, FOXO1, survivin, NF-κB and IκB. In NSCLC, selinexor forces nuclear retention of TSPs, inhibits tumor growth and NF-κB transcriptional activity, and induces cell death regardless of p53 status. These data demonstrate therapeutic potential for the treatment of NSCLC.

1

Authors: Gibson MK

Head and neck cancer (HNC) is the sixth most common cancer in the United States. Toxicity during treatment and disease itself may cause long-term disabilities in HNC patients and decrease quality of life (QoL). With the increasing numbers of HNC survivors it becomes even more important to address QoL issues.

4

Authors: Ali N

Background: Several mistakes in the diagnosis and treatment of bone tumors can be made, especially in non-specialized centers. Implanting conventional prostheses on an unrecognized bone tumor causes contamination of the entire region with dramatic consequences for prognosis. Purpose of our study was to try to understand which is the best way to deal with these patients. Does previous surgery affect prognosis? Does external hemipelvectomy achieve a better overall survival and local control than limb salvage surgery? Hypothesis: Demolitive surgery, scarifying the involved limb ensures better local control of the disease and improves life expectancy. Patients and Methods: We retrospectively evaluated all patients with bone sarcomas at the site of a total hip arthroplasty (THA) over the years 2000-2012. After reviewing the preoperative imaging and histological slides, 11patients had a THA implanted on an unrecognized hip sarcoma. Diagnosis was chondrosarcoma in 10 patients and osteosarcoma in one. Five patients were immediately treated with external hemipelvectomy. Results: Five of 11 patients (45%) died of disease at a mean time of 34 months (range 2-82 months), 4 are alive with disease and only 2are continuously disease free. Six of eleven patients (55%) had a local recurrence at a mean time of 17 months (range 3-36 months); six of these patients had conservative treatment. Conclusions: Although a very rare event, failure to recognize an occult malignant bone tumor during total hip arthroplasty associates with poor survival rate. Outcome after limb saving surgery is disappointing due to a high rate of local recurrences. According to our experience external hemipelvectomy provides better local control but this condition remains a dramatic event.

5

Authors: Abate SM

Purpose: The essence of this study is to illustrate the trends of breast cancer among women living in different regions of Ethiopia and to offer suggestions about some measures to put in place to control the disease and reduce its morbidity and mortality. Methods: Data were collected from October 2014 to April 2015. Data on new cases of breast cancer registered annually at the cancer registry of the Tikur Anbessa Specialize Hospital (TASH) over a period of sixteen years were obtained retrospectively and analysed using MS office and SPSS Version 20. Results: There were 3460 new cases of breast cancer registered at the cancer registry during the16-year period. The peak age of incidence was the 4th and 5th decade. Most of the cases were found in Addis Ababa, where the hospital is situated. An increase in trend of breast cancer case was observed in the hospital. Conclusion: Non-declining incidence of breast cancer in this study indicates; the awareness of people to be diagnosed is improved and more cancerous patients are there in the country, and inadequate or ineffective control measure to stem its morbidity due to diversion of the health care system’s attention to HIV/AIDS and malaria. Therefore, there is the need to step up activities through NGOs, screening programs and trainings such as mammography, clinical and self- examination, to control the upward trends of breast cancer in the country. In addition, it will be important to open breast cancer diagnosing centre in each region of the country in order to know the number of cases, which will be affected by being undiagnosed due to farness to the diagnosing centre. Additional researches on breast cancer post treatment survival rate, on risk factors assessment, patient quality of life and others are recommended in future.

2

Authors: Habip Gedik

Hyperammonemia syndrome is one of the causes of metabolic encephalopathy that is rarely observed after high dose chemotherapy for the treatment of hematologic malignancies. Two cases, who developed neurologic symptoms and coma due to hyperammonemia subsequent to chemotherapy for Burkitt’s lymphoma and acute myeloid leukemia, respectively are being presented in this report. In case respiratory alkalosis, unexplained neurological symptoms and mental status changes develop after intensive chemotherapy, hyperammonemia should come into mind, and the serum ammonium level should be measured to implement ammonium-lowering therapy.

5

Authors: Koul R

Introduction: Overall incidence rate of all brain tumors to be 10.82 (95% CI: 8.63–13.56) per 100,000 person-years. Glioblastoma accounts for up to 60% of all malignant primary brain tumors in adults, occurring in 2-3 cases per 100,000 in Europe and North America. In 2005 maximum safe surgical resection, followed by radiotherapy with concomitant temozolomide (TMZ), followed by adjuvant TMZ became the standard of care for glioblastoma. We adopted this as the standard of care in 2009 in the province of Saskatchewan, Canada. Material and methods: A cohort of 393 consecutive patients with pathologically proven glioblastoma, who had been registered in the Province of Saskatchewan from 2000 to 2010, was examined. Survival analysis was performed using Kaplan-Meier curves and log-rank test for comparing subgroups. The independent effect of factors that predicted survival at the bivariate level was determined using a Cox proportional hazard model. Results: Median age at diagnosis was 67 years in females and 63 years in males. The median overall survival was 13.8 months (95% CI: 12.6, 15.1 months). Based on a literature review and after the univariate analysis, the following variables were included in the Cox’s multivariable model: age at diagnosis, ECOG status (dichotomous variable created), type of surgery (complete vs. sub-total), and whether chemotherapy and radiotherapy were given after surgery. Patients who treated with chemotherapy and chemotherapy had a better median survival of 18.1 months vs. 11.3 months without chemotherapy. Patients younger than 50 years did better as compared to elderly population. For fit elderly patients >70 years, 11.0 months median survival was achieved. Contrary to common belief in literature, patients with headache do not have a worse survival and patients who presented with seizure survived better. Conclusion: Our series demonstrates improved survival outcomes for patients <50 years old quite consistent with literature. However for elderly patients with excellent performance status in the western Canadian province, a median survival of <70 years 16.1 months (95% CI 13.6- 18.1 months) ≥ 70 years 11.7 months (95% CI: 10.4-13.0 months) was achieved which is better than other series in the current literature. The landscape of treatment options for GBM patients has changed substantially over the past decade and with further information still amassing in ongoing clinical trials in GBM population the suggestion is to base treatment options based on patient age and KPS. Until further treatment advances are made for GBM in general, utilizing the current therapeutic options of surgery, RT, and TMZ appropriately according to patient age, performance status, and patient preferences represents optimal management.

8

Authors: Sita Aggarwal

Conjugates of membrane disrupting lytic peptides with a 15-amino acid segment of the β chain of chorionic gonadotropin (CG) or luteinizing hormone releasing hormone (LHRH) target and destroy cancer cell xenografts in nude mouse model. Follicle stimulating hormone receptors (FSHR) have been detected in tumor neo-vascular endothelial cells of a variety of cancers and tumor tissue in prostate and ovarian cancers. In the present study, we have conjugated a lytic peptide (Phor18) to each of three segments of the β chain of FSH that bind to FSHR, and tested these conjugates (FSH90-95-Phor18, FSH81-95-Phor18 and FSH33-53-Phor18) for their ability to target and inhibit growth of prostate cancer cells in vivo. We found that intravenous administration of FSH90-95-Phor18, FSH81-95-Phor18 and FSH33-53-Phor18 significantly (p<0.05) inhibited the growth of prostate cancer (PC-3) xenografts in nude mice. The minimal effective dose of FSH81-95-Phor18 was 0.1 mg/kg and 1 mg/kg for FSH90- 95-Phor18 and FSH33-53-Phor18. Immunohistochemical analyses of tumor sections at necropsy showed dense expression of FSHR on vascular endothelial cells in control mice, reduced expression in tumors of mice treated with FSH90-95-Phor18, FSH81-95-Phor18 or FSH33-53-Phor18. FSH81- 95-Phor18 had the most potent anti-angiogenic activity of the three conjugates tested. These data show that a lytic peptide conjugated to FSH β chain segments bind to FSHR and are capable of inhibiting prostate cell tumor growth by targeting and destroying endothelial cells that express FSH receptors.

5

Authors: Ellinor IB Peerschke

Background: gC1qR is a multifunctional cellular protein that has been linked to inflammation and cancer. gC1qR is highly upregulated in adenocarcinomas as compared to normal tissue counterparts, and soluble gC1qR (sgC1qR) has been detected in vitro in the pericellular milieu of proliferating malignant cells. Aim: The present study explored the tissue expression of gC1qR in pancreatic cancer by immunohistochemistry, and the presence of sgC1qR in vivo, by examining blood and malignant effusions from patients with metastatic pancreatic adenocarcinoma. Methods: Tissue expression of gC1qR by pancreatic adenocarcinoma was visualized by immunohistochemistry. SgC1qR was quantified in serum from healthy volunteers (n=20) and pancreatic cancer patients (n=34), as well as in malignant pleural (n=23) and peritoneal effusions (n=27), using a newly developed, sensitive immunocapture sandwich ELISA. Results: Overexpression of gC1qR was confirmed in pancreatic adenocarcinoma compared to nonmalignant pancreatic tissue. Moreover, increased serum levels of sgC1qR (0.29 + 0.22 ng/ml) were noted in patients with metastatic pancreatic cancer compared to healthy controls (0.15 + 0.10 ng/ml) (mean + S.D.) (p=0.035). In 11 of 16 patients for whom sequential samples were available, serum sgC1qR levels rose with disease progression, and paralleled changes in tumor biomarkers, CEA and CA19.9. In addition to blood, sgC1qR was detected in malignant pleural (0.55 + 0.47 ng/ml) and peritoneal effusions (0.57 + 0.38 ng/ml). Conclusion: This study provides the first evidence for the presence of sgC1qR in vivo. The ability to detect sgC1qR in blood and body fluids will enable further studies to elucidate its pathophysiology in malignancy.

6

Authors: Paul B. Tchounwou

Cis-diamminedichloroplatinum (II) (cisplatin) is the most widely used chemotherapeutic drug for various cancers, but its effectiveness is limited by tumor cell resistance and the severe side effects it causes. Since high level of cisplatin is cytotoxic to both cancer and normal cells, the goal of the present study was to explore the effectiveness of prolonged low doses of cisplatin in the management of leukemia. To achieve our goal, human leukemia (HL-60) cells were treated with different doses (1, 2, or 3 µM) of cisplatin for 24, 48, 72 and 96 hours. Cell viability was assessed by MTS assay. Both oxidative stress damage and genotoxicity were estimated by antioxidants, lipid peroxidation, and comet assays, respectively. Data obtained from the MTS assay demonstrated that cisplatin treatment decreased the number of viable tumor cells by direct cell killing or by simply decreasing the rate of cellular proliferation in a dose- and time-dependent fashion. The results of the lipid peroxidation showed a significant increase (p<0.05) of malondialdehyde levels with increasing cisplatin doses. Results obtained from super oxide dismutase and catalase assays showed a gradual increase in antioxidant enzyme activity in cisplatin-treated cells compared to control cells. Data generated from the Comet assay demonstrated a significant dose-dependent increase in genotoxicity with respect to DNA damage as a result of cisplatin treatment. Taken together, our research demonstrated that cisplatin-induced cytotoxicity in HL-60 cells is mediated at least in part via induction of oxidative stress and oxidative damage.

8

Authors: Susan Kadlubar

Anastrozole is an aromatase inhibitor (AI) used as adjuvant therapy for breast cancer. Anastrozole is subject to direct glucuronidation catalyzed by UDP-glucuronosyltransferase1A4 (UGT1A4). Interindividual variability in anastrozole glucuronidation may be affected by UGT1A4 SNPs. Interplay between drug metabolizing genes such as UGT1A4 and transporter genes may also be affected by genetic variability. Thus, we hypothesize that genetic variability in MRPs could influence anastrozole glucuronidation. The correlation between UGT1A4 and MRP2 or MRP3 transporter gene expressions and the correlation between MRP2 or MRP3 mRNA and anastrozole glucuronidation were analyzed in normal human liver samples. MRP2 and MRP3 mRNA levels were significantly correlated with UGT1A4 mRNA, with anastrozole glucuronidation and with each other (p<0.05). The data also demonstrated that MRP2 SNPs are positively correlated with MRP2 mRNA expression, while there was no association between MRP3 SNPs from this study and MRP3 expression. Significant correlations (p<0.05) between certain MRP2 SNPs (3972C>T, 2366C>T and -24C>T) and anastrozole glucuronidation were observed. There were no observed correlations between MRP3 SNPs and anastrozole glucuronidation. MRP2 polymorphisms have been identified as playing a role in the disposition of other drugs, and the data presented here indicate for the first time that MRP2 SNPs could influence anastrozole metabolism and contribute to interindividual variation in treatment responses.

3

Authors: Persinger MA

Aim: Discern if there is a specific proportion of mixture of normal and malignant cells that increase photon emissions from cell cultures. Method: Different proportions of B16-B6 mouse melanoma cells and HEX cells were mixed and allowed to proliferate. Photon emissions were measured from the different mixtures of cells by digital photomultiplier units and then analyzed for Spectral Power Density (SPD). Results: Mixtures of the malignant and non-malignant cells that were more than 10% of one component displayed photon emission flux densities that were significantly less than the photon emissions for either cell type when measured as pure samples. Only 1% proportion of the malignant cell in 99% of non-malignant cells produced the strongest photon emissions. Spectral density profiles of power flux density variations indicated elevated power around 22 Hz that was even greater than this signature for malignant cells only. Conclusion: Only 1% of malignant cells in a normal aggregate, representative of the early stages of cancer development, resulted in conspicuously increased numbers of photon emissions and spectral power spectra that often reflect a total malignant cell population. This combination of photon flux density and spectral power profiles may be a potentially useful (nanotechnology) tool to detect the minute changes in cell activities relevant to oncology.

5

Authors: Sergei V Jargin

After the Chernobyl accident appeared many publications exaggerating its medical consequences. Some of them are discussed in this minireview, which is a continuation of a preceding discussion. Among the motives for the overestimation were anti-nuclear resentments, widespread among the Green movement; however, their attitude has not been entirely wrong: nuclear facilities should have been prevented from spreading to overpopulated countries governed by unstable regimes and regions where conflicts and terrorism cannot be excluded. Nevertheless, we believe that certain Green activists have worked in the interests of the fossil fuel producers. The Chernobyl accident has been exploited to strangulate worldwide development of nuclear energy. Today, there are no alternatives to nuclear power: nonrenewable fossil fuels will probably become more expensive in the long run, contributing to the excessive population growth in the oil-producing countries and poverty in the rest of the world. Worldwide use of nuclear energy will become possible only after a concentration of authority within an efficient international executive based in the most developed countries. This will enable construction of nuclear power plants in optimally suitable places, considering all sociopolitical, geographic, geologic, and other preconditions, quality of working of local workforce, etc.

6

Authors: Jennifer Wu

Cholangiocarcinoma is a potentially lethal cancer of biliary epithelium with variable incidence rates across the world. Given patients often have advanced disease at the time of diagnosis and limited therapeutic options, the prognosis of patients with cholangiocarcinoma remains poor. There are efforts to develop new treatments for cholangiocarcinoma as current standard chemotherapy offers limited benefit. In vitro and in vivo models of cholangiocarcinoma have been studied to unveil underlying molecular mechanisms of cholangiocarcinogenesis. Recent advances in understanding of different pathways underlying cholangiocarcinogenesis will guide the development of potential therapeutic molecular targets. We will do a detailed review of the pathogenesis of cholangiocarcinoma and its relevance in molecular targets for potential therapies in cholangiocarcinoma. Clinical trials using targeted therapies and proposals for potential clinical trials using novel targets will be discussed.