Middle East Journal of Cancer
Volume:11 Issue: 4, Oct 2020

MicroRNAs (miRNAs) are short non-protein coding and single-stranded small RNA molecules with a critical role in the regulation of gene expression. These molecules are crucial regulatory elements in diverse biological processes such as apoptosis, development, and progression. miRNA genes have been associated with various human diseases, particularly cancer, and considered as a new biomarker. After the discovery of miRNAs, many researches have focused on identifying and characterizing miRNA genes in cancer. The various expression levels of miRNAs between cancer cells and normal cells are very crucial to diagnosis, prognosis, and treatment of many cancers. Many computational and experimental tools have been employed to characterize miRNAs. However, there exist some challenges in identifying miRNA using both computational and experimental tools due to miRNA features. The present review briefly introduced miRNA biology and certain computational and experimental tools for identifying and profiling miRNAs in cancer. Furthermore, we presented the advantages and challenges of these tools.

Oral squamous cell carcinoma (OSCC) is the sixth most common cancer worldwide and has a poor prognosis. The breast cancer 1 (BRCA1) and breast cancer 2 (BRCA2) genes are the key tumor suppressor genes responding in the cases of DNA damage. They repair double-strand DNA breaks to maintain gene stability. Mutations in BRCA1 and BRCA2 lead to genetic instability and develop different cancers, mainly familial breast and ovarian cancers. This study aimed to investigate the expression profiles of BRCA1 and BRCA2 genes in OSCC through the use of immunohistochemistry (IHC) technique. In this retrospective study, a total of 60 samples (20 samples of each grade) were collected from the archive of pathology department of Taleghani educational hospital, Tehran, Iran, from 2000-2017. IHC staining was performed for all tissue samples. BRCA1 immunoreactivity was positive in the cytoplasm and nuclear of 56 and 24 samples, respectively. None of the cancer cells showed nuclear BRCA2 expression; however, BRCA2 cytoplasmic staining existed in 17 cases. Chi-square test showed statistically significant differences between BRCA1 staining (P=0.001) and histological grade, and between BRCA2 expression (P=0.001) and histological grade in the research groups. Altered subcellular localization of BRCA1/2 and immunostaining of the cancer cells at the invasive front may indicate the critical role of BRCA1/2 in the development of OSCC. Early detection of BRCA mutation carriers by IHC has a significant impact on successful treatment.

The classification of genetic variations depending on their clinical impacts is highly relevant for clinical decision-making. Therefore, predicting the effects of missense mutations using in silico tools has become a frequently employed approach. The objective of this study was to analyze the impacts of a previously detected BRCA1 missense mutation using an in silico prediction tool in the context of invasive breast cancer. In this bioinformatics study, application of the in silico combination of tools Phyre2 to 184(T>C), a BRCA1 missense mutation previously characterized by Khalil et al. in 2018, we used to predict its clinical impacts. Incidence of the missense mutation caused a disorder in the zinc binding RING finger functional domain of BRCA1 protein. This incidence was considered to be a major contributor to the interaction of this protein with other proteins in signal transduction pathway in the mechanism of cellular response to DNA damage. The functional analysis also revealed that the detected missense mutation might significantly affect the function of both the protein and phenotype of the living organism. In silico prediction confirmed the detrimental impact of the identified missense mutation in the exon 2 of BRCA1 gene on both the structural and functional properties of the generated BRCA1 protein.

Aberrant expression level of Hox transcript antisense intergenic RNA (HOTAIR) has been associated with the etiopathogenesis of numerous cancers. Studies on epidemiological data have demonstrated that the risk of susceptibility to colon cancer varies among different populations due to several reasons. In this study, we aimed to assess the expression level of HOTAIR in tumoral tissues of patients with colon cancer and compare it with normal marginal tissues.

In this case-control study, we recruited a total of 50 patients with colon cancer and collected tumoral and matched marginal tumor free tissues during surgery. Afterwards, we isolated the total RNA from each sample, synthesized cDNA, and performed quantitative analysis by Real-time PCR using the SYBR Green PCR Master Mix in order to measure the transcript level of HOTAIR in samples.

The expression level of HOTAIR was upregulated in tumor tissues compared with normal tumor-free marginal tissues belonging to colon cancer patients (P= 0.0023). Moreover, the expression level of HOTAIR and the clinicopathological specifications of the patients had statistically significant correlations.

HOTAIR may play a role in the development of colon cancer and have the potential for application as a biomarker for colon cancer prognosis.

Cancer/testis antigens are a unique class of tumor antigens with normal expressions restricted to the testis and various cancers, but not in adult somatic tissues. Sperm-associated antigen 9 (SPAG9) has been introduced as a new member of Cancer Testis Antigens family involved in c-Jun-NH2-kinase signaling module. The objective of this research was to investigate the potential of SPAG9 as a diagnostic and prognostic biomarker in breast cancer. We further aimed to find any significant association between SPAG9 expression and clinicopathologic features of the cancer.
In this retrospective study, 35 breast cancer tissues and 35 adjacent non-cancerous tissues were collected and examined using RT-PCR to explore SPAG9 mRNA expression. Statistical analysis was done utilizing SPSS 22.0 software.
Unexpectedly, we detected SPAG9 expression in 54% of adjacent non-cancerous tissues. Moreover, SPAG9 mRNA was expressed in 57% of cancerous tissues. Statistical analysis showed a significant association between SPAG9 expression and tumor size, lymph node metastasis, and cancer stage.
The association between the gene expression and tumor size, lymph, node and metastasis, and cancer stage suggests that SPAG9 can potentially be considered as a prognostic biomarker in breast cancer. However, it may not be a candidate diagnostic biomarker.

The preponderance of breast cancer-related deaths are the result of local invasion and distant metastasis; therefore, it is necessary to identify the factors underlying invasion and metastasis in order to develop novel treatment strategies and improve the survival of patients. In this regard, this study aimed to investigate the immunohistochemical expression and prognostic impact of aquaporin-3 (AQP3) and certain markers associated with epithelial-mesenchymal transition concerning invasive breast carcinoma of no special type.

Immunohistochemical expressions of AQP3, vimentin and E-cadherin were performed in 50 paraffin embedded specimens of such cases. We also assessed the relationship of their expressions with the clinicopathological variables and patients’ disease-free survival and overall survival.

There were significant associations between positive AQP3 and positive vimentin expressions and high tumor grade, large tumor size, lymph node metastasis, and advanced tumor stage. On other hand, negative E-cadherin expression had a significant correlation with high tumor grade, large tumor size, lymph node metastasis, distant metastasis, and advanced tumor stage. A significant association also existed between positive AQP3, positive vimentin and negative E-cadherin expressions and high tumor recurrence, short ‘three-year’ disease-free survival and overall survival.

Positive AQP3, positive vimentin, and negative E-cadherin expressions are known as adverse prognostic markers and may predict survival in invasive breast carcinoma of no special type. It is proposed that AQP3 might play a role in breast cancer progression, invasion, and metastasis through induction of epithelial-mesenchymal transition.

Human epididymis protein 4 (HE4) is a novel tumor marker that has shown a strong potential in diagnosing and predicting the prognosis of endometrial cancer. In this study, we aimed to assess the association of tumor size and stage, ovarian involvement, lymph node metastasis, lymphovascular space invasion and deep myometrial invasion with preoperative serum levels of HE4 in endometrial cancer patients. This cross-sectional study included patients with endometrial cancer awaiting surgery in Ghaem Hospital, Mashhad, Iran, from May 2016 to May 2017. We measured the HE4 levels preoperatively and gathered other required information postoperatively; collected data were analyzed using SPSS version 16. We enrolled 104 endometrial cancer patients. Mean serum HE4 was 149.6±186.2pmol/l. Level of serum HE4 had significant positive correlations with tumor size and stage, lymph node metastasis, ovary involvement, lymphovascular space invasion and profound myometrial invasion (p <0.001). There was no association between age and myometrial invasion. Moreover, using ROC curve, we calculated a serum HE4 cut-off of 104 pmol/l to be 91% sensitive and 89.6% specific for the detection of deep myometrial invasion. HE4 is a novel biomarker capable of preoperatively predicting the depth of myometrial invasion with high specificity and sensitivity. This marker can be utilized in guiding the surgical plan of patients with endometrial cancer.

The objective of this study was to investigate the effect of several XPA and XPC polymorphisms on the risk of colorectal cancer (CRC) in northeastern Iran.

180 CRC patients and 160 healthy subjects participated in this case-control study. We determined the genotypes by RFLP-PCR and PIRA-PCR, and analyzed the results using logistic regression and χ2-test.

Our findings showed that only BMI could affect the risk of cancer among the studied demographic factors. Three of the four polymorphisms studied, namely XPA A23G, XPC rs2228000 C > T and XPC rs2228001 A > C, did not correlate with CRC (P-values > 0.05); however, the polymorphism of XPC poly AT (PAT) increased the risk of CRC (P= 0.024). The XPC rs2228000 C> T polymorphism increased the CRC risk only in patients aged 50 or more. The risk of CRC in heterozygote individuals (XPC PAT D/I) was higher than that of homozygous individuals (XPC PAT D/D); also, at least one PAT I variant allele increased the likelihood of CRC (for PAT D/I OR =2.168; 95% CI = 1.809-4.319: and for PAT D/I and PAT I/I OR = 1.810; 95% CI = 1.165-2.813). The XPC haplotypes were similar between the cases and controls, and P-values were >0.05.

In the whole population, XPC PAT polymorphism, overweightness, and XPC rs2228000 C>T polymorphism in elderly people are related to CRC. Therefore, they can probably be considered as markers of CRC in Iran.

The incidence rate of cervical cancer is increasing and its existing drugs are becoming more and more resistant. Therefore, we extracted the fruiting body of Calocybe indica edible mushroom in 90% ethyl acetate extract (EAE) and evaluated it as an anticancer property against HeLa and CaSki.
We performed cytotoxicity assay by MTT, cell morphological study by phase contrast microscope, and apoptosis study by nuclear morphology via DAPI staining under inverted microscopy; the expressions of proapoptotic and antiapoptotic genes and p53 were examined by Western blotting, cell cycle analysis, and cologenic and cell migration assay. Antioxidant content and activity assays were performed and for mycochemistry analysis of EAE, thin layer chromatography (TLC) was done.
EAE-treated HeLa and CaSki cells became round and showed condensed and fragmented nuclei. They inhibited the cell proliferation of both cancer cell lines in a dose-dependent manner. At maximum dose (1250 μg/mL) after 24 h, the cell inhibition percentages of HeLa and CaSki cells were 97.12±10.01 and 98.52±10.08 (P<0.05), respectively. They upregulated the expression of p53, caspase 3, and caspase 9 while down-regulating BcL2 gene. Cell cycle became arrested at G2/M checkpoint of both cancer cell lines by EAE. EAE inhibited colony formation and cell migration. The antioxidant assay showed that EAE contained good amounts of phenolic compounds, flavonoids, and ascorbic acids and had good antioxidant activity. TLC supported the presence of bioactive components.
The EAE of C. indica exerts very potent anticervical cancer effects. It is urgent that future studies analyze its bioactive compounds in detail and examine them in animal models.

T-cell acute lymphoblastic leukemia is an aggressive hematologic malignancy that results from the transformation of T-cell progenitors. Despite the significant advances in the current treatments, the side-effects of conventional chemotherapy regimens are still a major concern. Pterostilbene (PT) is a natural compound reported to have antitumor effects. This study aimed to investigate the effect of PT combined with dexamethasone on the proliferation inhibition and apoptosis stimulation in a lymphoblastic leukemia cell line.

In this experimental study, we cultured Jurkat cell line in RPMI1640 culture medium under standard conditions. We incubated the cells with different concentrations of PT and dexamethasone, separately or in combination for 48 h. MTS assay investigated the cell viability. We assessed apoptosis induction by annexin V-FITC/PI and flow cytometry analysis.

PT and dexamethasone reduced the viability of the cells with inhibition concentrations of 60.97±3.36 and 451.1±10.1 μM, respectively, in 48 h. None of the concentrations of dexamethasone, employed alone, significantly reduced the cell viability. The combination of 450 μM dexamethasone with 60 μM PT induced apoptosis in more than 70% of the cells with a significant difference compared to control.

PT increased the antiproliferative and apoptosis-inducing activity of dexamethasone in Jurkat cell line. This combination drug strategy can be a novel approach for a more powerful anticancer therapy.

Synovial sarcoma is an aggressive soft tissue sarcoma. It has a wide spectrum of histopathologic patterns and uncertain immunohistochemistry, rendering it a diagnostic challenge. The objective of this study was to investigate the diagnostic utility and impact of SS18-SSX rearrangement evaluation in an Iranian population previously diagnosed with synovial sarcoma without such molecular tests.

We conducted this cross-sectional study on 44 formalin-fixed paraffin-embedded tissue blocks obtained from 23 synovial sarcoma patients (males, 69%; mean age, 36.4 years) and 11 cases with other neoplasms as negative controls. We assessed these specimens for SSX-SS18 gene rearrangement by break-apart fluorescent in situ hybridization (FISH) probes.

FISH study showed SS18-SSX fusion gene in 17 (73.91%) cases while six (26.09%) cases and negative controls did not show SS18-SSX fusion. Histopathologic type of tumor was significantly related to the presence of rearrangement (P=0.002) (rearranged: 11 biphasic and six monophasic tumors; non-rearranged: three monophasic and three poorly-differentiated tumors).

This study supports the idea that molecular studies contribute to the confirmation of synovial sarcoma diagnosis, particularly in monophasic and poorly-differentiated subtypes, which show vague immunohistochemical results.

Colorectal cancer (CRC), caused by abnormal cells growing in the colon or rectum, has a high mortality rate worldwide. On the other hand, microRNAs are small non-coding RNAs that contain approximately 22 nucleotides in length. They are upregulated in a wide range of human cancers such as CRC. MiRNA-21 post-transcriptionally regulates the expression of many tumor suppressor genes such as P53 gene. This indicates that miRNA-21 interacts like oncogenes and is required for CRC development.
The current original study was conducted in the National Liver Institute, Menofyia University, Egypt. We collected a total of 40 blood samples from CRC patients 40 samples from healthy individuals who served as controls. Quantitative real-time PCR detected the levels of miRNA-21 and the fold changes of phosphates-tensin homology (PTEN) gene expression, as a tumor suppressor gene, in blood samples.
The expression levels of miR-21 were upregulated in all obtained samples from patients with CRC in association with aging, gender, and tumor-node-metastasis staging. Furthermore, patients with poor and well-differentiated CRC revealed reduced levels of PTEN gene expression. We observed a putative binding site of miR-21 in PTEN gene sequences. This indicates the direct cleavage between miR-21 and PTEN coding sequence. Prediction analysis for other potential targets identified several malignancy factors and tumor suppressor genes with putative seeding regions for miR-21 such as STAT3, transforming growth factor-beta, tumor necrosis factor-α (TNF-α), and programmed cell death CD4.
The current data exhibited the potential dual role of hsa-miR-21 in regulating cancer progression and showed that hsa-miR-21 is an efficacious biomarker for CRC d evelopment and an attractive candidate for CRC treatment during early transformation.

Hereditary diffuse gastric cancer (HDGC) is a hereditable form of diffuse gastric cancer with very aggressive tumors, poor prognosis, and delayed clinical signs.

We assessed 17 probands identified with HDGC upon gastrectomy according to the histopathological criteria confirmed by a pathologist and familial history. We extracted DNA from peripheral blood and formalin fixed paraffin-embedded tissues. DNA sequencing was done following PCR amplification of 16 exons and exon/intron boundaries of the CDH1 gene and exon 2 of CTNNA1 gene. The Multiplex Ligation-dependent Probe Amplification technique was performed on patients with no pathogenic variants in sequencing.

Totally, 17 probands comprising seven males and 10 females were assessed. In three patients, we recognized the tumors in the early TNM stage (I, II), while in 14 cases, tumors were observed in the late stages (III, IV). Overall, DNA sequencing of the CDH1 gene identified 16 variants (seven exonic including five new variants and nine intronic containing six new variants). Moreover, Multiplex Ligation-dependent Probe Amplification detected one deletion in exon 1 of two patients.

Our results showed that E-cadherin deficiency in HDGC was related to CDH1 gene point mutations and large deletion with high heterogeneity, which should be considered in the diagnosis and treatment of HDGC patients.

Hematopoietic stem cell transplantation is a potentially life-saving therapy under certain conditions. Hypersensitivity reaction to chemotherapeutic drugs may interfere with this treatment. Griscelli syndrome type 2 is a rare autosomal recessive disease characterized by hypopigmentation of skin and hair, causing silver gray hair; this disease may develop a fatal condition called hemophagocytic lymphohistiocytosi. The main curative treatment for hemophagocytic lymphohistiocytosi is hematopoietic stem cell transplantation. Melphalan is an important drug in hematopoietic stem cell transplantation preparation regimen. This drug is only stable for 60 minutes following reconstitution with normal saline. There is a recommended 12- to 20-step desensitization protocol for moderate to severe hypersensitivity reaction to chemotherapeutic drugs. However, due to the short term stability of melphalan, this protocol cannot be employed in desensitization. In this case report, we presented a novel protocol for rapid desensitization of melphalan in a 5-year-old boy with immediate, moderate to severe hypersensitivity reaction, during hematopoietic stem cell transplantation. To obtain 50 mg total melphalan , we prepared solutions of drug in two bags; one with 12.5mg in 35 cc normal saline, and other with 25 mg in 35 cc normal saline. Each solution was infused during 60 minutes by incremental infusion rate, and the total of 50 mg melphalan was successfully infused.

We report a case of colon cancer with pancreatic and interesting heart metastasis several months prior to disseminated metastases. The case was a 59-year-old man with a cecal cancer (T4N2); he received chemotherapy with XELOX regimen and then radiotherapy up to a dose of 45 Gy. He was under close and regular follow-up. After 42 months, he developed jaundice and computed tomography (CT) scan showed an isolated mass in the pancreas. We performed Whipple’s operation and the pathology report was pancreatic metastasis. He received chemotherapy and was relatively well until his CEA rose again and the chest CT scan indicated cardiac metastasis. We resected the metastasis and administered chemotherapy. Unfortunately, the case developed brain metastasis and passed away. We searched the literature and found 15 cases of colon cancer and cardiac metastasis. We found no cases with metastasis from colon cancer to the left side. Although cardiac metastasis has a poor prognosis, it might be more prevalent than what is generally believed.

Acute lymphoblastic leukemia has several presentations associated with bone marrow and extramedullary involvement. The unusual presentation may be due to the infiltration of leukemic cells in any organ. An 11-year-old girl presented with fever and vomiting, since one day before admission after starfish biting during swimming. Her vital signs were: blood pressure 150/100 mmHg, pulse 98 beats per minute, respiration 18 breathes per minute, and temperature 37.2 °C (99 F). Laboratory work-up showed blood urea nitrogen 38 mg/dl and creatinine 2.8 mg/dl. In peripheral blood smear, few atypical cells, mild anemia (Hb: 9.2 g/dl), and mild thrombocytopenia (Platelet: 109,000/μL) were detected. Bone marrow aspiration and immunophenotyping were in favor of acute precursor B cell type lymphoblastic leukemia. The patient had a favorable response to treatment after initiating high-risk chemotherapy. Therefore, acute renal failure can be a rare initial presentation of acute lymphoblastic leukemia, and azotemia will improve with an early chemotherapy treatment.