Inhibition of breast cancer cell growth and migration through siRNA-mediated modulation of circ_0009910/miR-145-5p/MUC1 axis.

Circular RNAs (circRNAs) characterize a novel kind of regulatory RNAs distinguished by great evolutionary conservation and constancy. Although their exact role in malignancies is not fully understood, they mainly work through specific axes. Circular RNA/miRNA/mRNA axes affect the pathogenesis of human cancers including breast cancer. We assessed the expression and function of circ_0009910/miR-145-5p/MUC1 axis in Breast Cancer tissues and MCF-7 cells. Expression levels of circ_0009910 and MUC1 were notably increased in breast cancer tissues compared with control tissues, parallel with the down-regulation of miR-145-5p. Clinicopathological analysis indicated that up-regulation of circ_0009910 in breast tumors is related to invasion of the tumor to lymph node (P value = 0.011). Also, the downregulation of miR-145-5p was significantly correlated with tumor invasion to lymph nodes (P value = 0.04) and HER2-negative tumors (P value = 0.037). Finally, overexpression of MUC1 was correlated with age under 45 years (P value = 0.002). More importantly, circ_0009910-siRNA decreased the proliferation and migration ability of breast cancer cells, enhanced expression of miR-145-5p, and decreased levels of MUC1. Taken together, the circ_0009910/miR-145-5p/MUC1 axis has been demonstrated to affect the pathogenesis of breast cancer and might provide a target for breast cancer treatment.

LINC02381 suppresses cell proliferation and promotes apoptosis via attenuating IGF1R/PI3K/AKT signaling pathway in breast cancer.

Identification of the genes and genetic networks involved in breast cancer development is a major need for prevention and therapy. LINC02381 (lncRNA) has already been introduced as a tumor suppressor in colorectal and gastric cancers. Here, we intended to investigate its potential functional effects on breast cancer. In the analysis performed on RNA-Seq and microarray data, the LINC02381 lncRNA was found to be significantly downregulated in the breast tumors and associated with poor survival of the patients. Then, the differential expression of LINC02381 was confirmed in breast tumor tissues and cancer cell lines using RT-qPCR. Overexpression of LINC02381 resulted in reduced IGF1R and p-AKT expression levels which indicates decreased PI3K pathway activity, detected by RT-qPCR and western blotting. At the cellular level, LINC02381 overexpression was followed by a decreased proliferation rate of transfected breast cell lines, detected by PI flow cytometry, RT-qPCR, colony formation, and MTT assays. Consistently, the results of Annexin-V/PI flow cytometry, RT-qPCR, caspase3/7 activity, and AO/EB-H33342/PI dual staining revealed that LINC02381 overexpression induced apoptosis and cell death. The reduced migration rate of these cells was also verified through wound healing assay and RT-qPCR against the EMT-involved genes. Our data show that LINC02381 exerts its tumor suppressor effect at least partly through attenuation of the IGF1R/PI3K/AKT signaling pathway, which originated from IGF1R downregulation.

DOK7 CpG hypermethylation in blood leukocytes as an epigenetic biomarker for acquired tamoxifen resistant in breast cancer.

BACKGROUND: Breast cancer (BC) is among the most common cause of cancer 10.4% and one of the leading causes of death among 20-50 years old women in the world. Tamoxifen drug is the first line therapy for BC however tamoxifen resistance (TR) has shown in 30-50% of cases that may face BC recurrence. Hence, TR early detection reduces BC recurrence and fatalities. The epigenetic alteration that happens by hypermethylation of tumor suppressor genes and hypomethylation of oncogenes has been suggested to be useful in early cancer or drug resistance diagnosis. METHODS: This is the first study to investigate DOK7 CpG hypermethylation in blood leukocytes of 31 TR (ER+) BC compared to 29 tamoxifen sensitive BC to evaluate DOK7 as a potential TR biomarker. DNA was extracted from blood samples of all participants and MSRE-PCR and real-time PCR were used for quantification of CpG methylation alterations. RESULTS: The means of DOK7 CpG hypermethylation were obtained as 85.03%, 29.1% and 57.34% in TR, TS and normal control respectively. Significant hypermethylation were found among TR vs. TS (p < 0.001), TS vs. normal (p < 0.001) and TR vs. normal controls (p < 0.03). Online databases expression and survival analysis of DOK7 showed increasing expression in TS groups vs. TR groups which have consistency with our methylation alteration results. The sensitivity and specificity of the TR epigenetic test were determined using ROC analysis showed 89.66% and 96.77% respectively and showed that 37.5% above hypermethylation is at risk for TR and breast cancer recurrence. CONCLUSION: There is a significant difference in the methylation ratio of DOK7 between tamoxifen resistant and tamoxifen sensitive groups that may be useful in the early diagnosis of tamoxifen resistance in BC cases and cancer recurrence prevention.

Association between Oxidative Stress Parameters and Hematological Indices in Breast Cancer Patients.

Background: Breast cancer is one of the leading causes of death in women worldwide. This causes an increase in free radicals, resulting in oxidative stress. The aim of this study was to determine the effect of breast cancer on oxidative stress and its relationship with hematological indices. Methods: This case-control study included 43 women with breast cancer and 37 age-matched healthy controls. Oxidative stress and its correlation with hematological profiles over seven months were evaluated. Finally, the data were compared between the two groups using the t-test and Pearson's test, and the results were analyzed using the SPSS 24 software. Results: The results revealed that patients with breast cancer had significantly increased hemoglobin (HB), hematocrit (HCT), mean corpuscular volume (MCV), and mean corpuscular hemoglobin (MCH) levels compared with healthy subjects (p < 0.05). In addition, oxidative stress parameters, such as superoxide dismutase (SOD), catalase (CAT), total oxidant status (TOS), and total antioxidant capacity (TAC), were significantly elevated. Glutathione peroxidase (GPX) and malondialdehyde (MDA) were significantly lower in patients with breast cancer than in the control group (p < 0.05). Statistical significance in hematological indices showed a positive or negative correlation with oxidative stress parameters. Conclusion: Women with breast cancer showed a deranged complete blood count (CBC) pattern compared to healthy individuals.

Long Non-Coding RNAs Expression in Breast Cancer: CBR3-AS1 LncRNA as a Sensitive Biomarker.

BACKGROUND: Long non-coding RNAs (LncRNAs) are eminent genes in the human genome that interfere with the regulation of many complexities of organisms and control many of the various biological processes. As a result, it is considered that they may play an important role in different cancers. With regard to the high prevalence of breast cancer and the role of lncRNA, the present study aimed at investigating the expression of various lncRNAs. METHOD: Fresh tissues were obtained from operating rooms of Shariati, Khatamolanbia, and Milad Hospitals (Tehran, Iran) by a surgeon. A total of 45 tumor samples and 45 non-tumor samples (from the margin of tumor) were obtained from the same patients. Relative expression evaluation method was used in Real time PCR. Estrogenn receptor (ER), progesterone receptor (PR), and HER2 expression were analyzed using IHC analyses of each cell block. RESULTS: Participants included 44 female and 1 male with the mean age ± SD of 50 ± 12.0 years (range: 23-74). A majority of participants (41/45) were Ductal carcinoma type. Our results showed significant expressions for CBR3-AS1 (P-value=0.0139), RAB6C-AS1 (P-value=0.0023), and ZEB2-AS1 (P-value=0.0289) in comparison with the healthy cells. ROC curve analysis for CBR3-AS1 LncRNA revaled sensitivity more than 70%. CONCLUSION: Although CBR3-AS1, RAB6C-AS1, and ZEB2-AS1 lncRNAs were found to have high expressions in the breast cancer cells, only CBR3-AS1 lncRNA has a high chance to be a breast cancer biomarker.
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Serum Ferritin, Vitamin D and Pathological Factors in Breast Cancer Patients.

Background: Breast cancer is one of the most common cancers. Researchers are trying to diagnose the disease through easier and safer methods. Serum markers such as ferritin and vitamin D level would be very helpful. This research could pave the way for more comprehensive studies on how to use this serum factor in breast cancer screening, as well as early detection of the disease in its early stages. Methods: This study consisted of two groups, the first group comprising patients diagnosed with breast cancer before undergoing any treatment and the second group as control were healthy people. Serum ferritin and vitamin D levels were measured. Pathological information of the patient's tumor, including ER, HER2, KI67, lymphovascular invasion, and disease stage, were collected as well. Data were analyzed by IBM SPSS advanced statistics version 23.0 (SPSS Inc., Chicago, IL). P-value of ≤0.05 was considered significant. Results: Eighty-eight subjects were enrolled in this study, 29 (33%) breast cancer patients and 59 (67%) healthy women. In breast cancer patients, serum ferritin levels were 106.55±111.25, which were higher than healthy women's serum ferritin 52.71±36.95 (p=0.083). Furthermore, 18 (66.7%) of breast cancer patients and 55 (93.2%) of healthy women had low serum ferritin levels (p=0.001). 3 (11.1%) patients in the cancer group had serum vitamin D deficiency, while all subjects in the control group had serum vitamin D higher than 10 ng/dl (p =0.009). Conclusion: The results of this study showed a correlation between breast cancer and vitamin D deficiency, and elevated ferritin. Perhaps with further studies, there could be a role in predicting the prognosis and screening of breast cancer for these associations.

Sentinel Node Biopsy in Early Breast Cancer Patients with Palpable Axillary Node.

BACKGROUND: Sentinel lymph node biopsy is a reliable method for evaluation of the axillary lymph node status in early stage breast cancer patients with non-palpable lymph nodes. The present study evaluated the status of sentinel and non-sentinel lymph nodes in T1T2 patients with palpable axillary lymph nodes. MATERIALS AND METHODS: One hundred and two women with early breast cancer were investigated in this study. Patients were selected for axillary sentinel lymph node biopsy and then surgery .Then the rates of false negative and true positive, and diagnostic accuracy of sentinel lymph nodes biopsy were evaluated. In addition, the hormone receptors status of the tumor was determined through IHC and data was analyzed in SPSS21. RESULTS: In this study, the mean age of the patients was 49 years, 85% had invasive ductal carcinoma  in their pathology reports, 77% were ER/PR positive, 30% HER2 positive and 9.8% triple negative and 69% had KI67<14%. In frozen pathology, 15.7 and 84.3% were sentinel positive and negative, respectively, and in the final pathology, 41 and 58.8% were sentinel positive and negative, respectively. This difference arises from the false negative rate of the frozen pathology, which was about 31.3%. The sensitivity, specificity, and diagnostic accuracy of the frozen section were 24, 90 and 43%, respectively. Lymphovascular invasion is an important effective factor in the involvement of sentinel and non-sentinel lymph nodes. Statistical analysis showed that the probability of sentinel and non-sentinel lymph nodes involvement was higher in receptor positive patients and those with KI67>14% (p<0.002) whereas the rate of involvement was lower in triple negative patients. CONCLUSION: Sentinel node biopsy can be used in a significant percentage of breast cancer patients with palpable and reactive axillary lymph nodes.

Transcriptomic and proteomic approaches reveal biological basis of intraoperative radiotherapy-treated tumor bed modification in breast cancer patients: A pilot study.

AIM: Intraoperative electron Radiotherapy, herein referred to, as IOeRT is a novel approach in breast cancer (BC) treatment. This study designed to investigate short-term molecular effects of 12Gy as Boost versus 21Gy as Radical dose of IOeRT using high throughput approaches. MATERIALS AND METHODS: Six BC patients as a pilot study were treated with IOeRT following two separate strategies, including Boost and Radical doses. Approximately 100 mg of tumor bed tissue retrieved from each patient (before IOeRT,immediately, 24 h post-treatment). mRNA sequencing also Isobaric tag for relative and absolute quantitation (iTRAQ) were performed to study the transcriptome and proteome profile of IOeRT-treated tumor bed. RESULTS: Using NGS, ~6 Giga base (GB) clean data per individual samples were generated. Moreover, by iTRAQ for proteome quantification, in total, 1,045,410 spectrums were generated, likewise 5860 proteins were identified (FDR <0.01). CONCLUSION: Functional annotation and gene ontology (GO) indicated that significant enrichment in molecular pathways on BC treatment is somehow single high dose-independent. This means that, key molecular pathways in radiotherapy (RT) are equally enriched by both Boost and Radical doses. Generally, by modification of the Radical dose, with the same effectiveness, it is possible to reduce single high dose irradiation in BC.

Measurement of peripheral dose to the pelvic region and the associated risk for cancer development after breast intraoperative electron radiation therapy.

This study aimed to measure the received dose to the pelvic region of patients during breast intraoperative electron radiation therapy (IOERT). Furthermore, we compared the findings with those of external beam radiation therapy. Finally, secondary ovarian and uterus cancer risks following breast IOERT were estimated. In the current study, the received dose to the pelvic surface of 18 female patients during breast IOERT boosts were measured by thermoluminescent dosimeter (TLD-100) chips. All patients were treated with 12 Gy given in a single fraction. To estimate the dose to the ovary and uterus of the patients, conversion coefficients for depth from the surface dose were obtained in a Rando phantom. Given the received dose to the pelvic region of the patients, secondary ovarian and uterus cancer risks following breast IOERT were estimated. The received doses to the ovary and uterus surface of the patients were 0.260 ± 0.155 mGy to 31.460 ± 6.020 mGy and 0.485 ± 0.122 mGy to 22.387 ± 15.476 mGy, respectively. Corresponding intra-pelvic (ovary and uterus) regional doses were 0.012 ± 0.007 mGy to 1.479 ± 0.283 mGy and 0.027 ± 0.001 mGy to 1.164 ± 0.805 mGy, respectively. Findings demonstrated that the ratio of the received dose by the pelvic surface to the regional dose during breast IOERT was much less than external beam radiation therapy. The mean of the secondary cancer risks for the ovary in 8 and 10 MeV electron beam energies were 135.722 ± 117.331 × 10-6 and 69.958 ± 28.072 × 10-6, and for the uterus were 17.342 ± 10.583 × 10-6 and 2.971 ± 3.604 × 10-6, respectively. According to our findings, the use of breast IOERT in pregnant patients can be considered as a safe radiotherapeutic technique, because the received dose to the fetus was lower than 50 mGy. Furthermore, IOERT can efficiently reduce the unnecessary dose to the pelvic region and lowers the risk of secondary ovarian and uterus cancer following breast irradiation.

Ovotesticular Disorder of Sex Development Presenting as an Acute Scrotum.

Hermaphroditism is known as ovotesticular disorder of sex development. A 14-year-old boy was admitted with right acute scrotum. Exploration revealed tunica rupture and hematoma, with no viable tissue. After 1 month, he was admitted again with left hemiscrotal pain. Microscopic examination of the left gonad demonstrated foci of hemorrhagic cysts, primordial follicles, and regions of seminiferous tubules. We preserved a testicular tissue and the ovarian part was extracted completely. Long-term follow-up with his hormonal profile is reported. This is a case of ovotesticular disorder presented with acute scrotum and we also tried to reduce long-term hormone therapy, with preservation of testicular part.

The Relationship Between 6 Polymorphisms of Caveolin-1 Gene and the Risk of Breast Cancer.

BACKGROUND: Caveolae play a role in cell signal transduction, kinetic regulation of transport vesicles, and cellular physiology. In this study, we evaluated the role of caveolin-1 (CAV-1) genotypes in the risk of breast cancer. PATIENTS AND METHODS: We evaluated 6 single nucleotide polymorphisms of the CAV-1 gene in a sample size of 406 participants. Six polymorphisms-G32124A (rs3807992), T29107A (rs7804372), T28608A (rs3757733), G21985A (rs12672038), G14713A (rs3807987), and C521A (rs1997623)-were assessed using restriction fragment length polymorphism polymerase chain reaction. RESULTS: Regarding the distribution of genotypes, the relationship between cases and controls was significant for T29107A, G21985A, G14713A, and C521A polymorphisms, among which only C521A showed a significant difference in body mass index between the 2 groups. Moreover, the age of the 2 groups was significant in the case of G32124A and T28608A polymorphisms. CONCLUSION: Our results showed that genetic changes of CAV-1 might modify the risk for breast cancer and point out the importance of more studies for variants of this gene in breast cancer.

Application of Human Acellular Breast Dermal Matrix (ABDM) in Implant-Based Breast Reconstruction: An Experimental Study.

BACKGROUND: The use of acellular dermal matrices (ABDM) has become more common for breast reconstruction to improve postoperative outcomes. We evaluated the efficacy of breast reconstruction by the application of human ABDM in a sheep model. METHODS: The sheep in group I (GI) (N = 4) underwent the following procedures on the right side: (1) breast reconstruction using human ABDM after total mastectomy, (2) human ABDM under the skin, near the breast area and on the left side, (3) fat injection and human ABDM after partial mastectomy, and (4) replacement of ABDM in the abdominal wall far from the breast. Sheep in group II (GII) (N = 4) underwent the following procedures. On the right side: (1) breast reconstruction using ABDM after total mastectomy, (2) replacement of ABDM under the skin, near the breast area, and on the left side, (3) application of vicryl synthetic mesh after partial mastectomy and (4) replacement of mesh under the skin, near the breast area. RESULTS: Histological evaluations of decellularized skin scaffolds demonstrated a collagen-based matrix with preserved ECM and complete nuclear removal. Histological evaluations of implanted ABDM demonstrated a viable matrix with fibroblast infiltration and revascularization in all follow-ups. The overall surgical complication rate was significantly lower in the ABDM implant under the skin and near the breast in both short- and long-term follow-ups. CONCLUSION: The results of this study demonstrated that the application of novel prepared ABDMs has promising outcomes for breast reconstruction to provide total coverage without the need for breast expansion before implant placement. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors http://www.springer.com/00266.

Value of sentinel lymph node biopsy in breast cancer surgery with simple pathology facilities--an Iranian local experience with a review of potential causes of false negative results.

INTRODUCTION: Sentinel lymph node biopsy (SLNB) is a precise procedure for lymphatic staging in early breast cancer. In a valid SLNB procedure, axillary lymph node dissection (ALND) can be omitted in node- negative cases without compromising patient safety. In this study, detection rate, accuracy and false negative rate of SLNB for breast cancer was evaluated in a setting with simple modified conventional pathology facilities without any serial sectioning or immunohistochemistry. MATERIAL AND METHOD: Patients with confirmed breast cancer were enrolled in the study. SLNB and ALND were performed in all cases. Lymph node metastasis was evaluated in SLN and in nodes removed by ALND to determine the false negative rate. Pathologic assessment was carried out only by modified conventional technique with only 3 sections. Detection rate was determined either by lymphoscintigraphy or during surgery. RESULTS: 78 patients with 79 breast units were evaluated. SLN was detected in 75 of 79 cases (95%) in lymphoscintigraphy and 76 of 79 cases (96%) during surgery. SLN metastases was detected in 30 of 75 (40%) cases either in SLNB and ALND groups. Accuracy of SLNB method for detecting LN metastases was 92%. False negative rate was 3 of 30 of positive cases: 10%. In 7 of 10 cases with axillary lymphadenopathy, LN metastastates was detected. CONCLUSION: SLNB is recommended for patients with various tumor sizes without palpable lymph nodes. In modified conventional pathologic examination of SLNs, at least macrometastases and some micrometastases could be detected similar to ALND. Consequently, ALND could be omitted in node-negative cases with removal of all palpable LNs. We conclude that SLNB, as one of the most important developments in breast cancer surgery, could be expanded even in areas without sophisticated pathology facilities.