Dynamically decreased miR-671-5p expression is associated with oncogenic transformation and radiochemoresistance in breast cancer.

BACKGROUND: Understanding the molecular alterations associated with breast cancer (BC) progression may lead to more effective strategies for both prevention and management. The current model of BC progression suggests a linear, multistep process from normal epithelial to atypical ductal hyperplasia (ADH), to ductal carcinoma in situ (DCIS), and then invasive ductal carcinoma (IDC). Up to 20% ADH and 40% DCIS lesions progress to invasive BC if left untreated. Deciphering the molecular mechanisms during BC progression is therefore crucial to prevent over- or under-treatment. Our previous work demonstrated that miR-671-5p serves as a tumor suppressor by targeting Forkhead box protein M1 (FOXM1)-mediated epithelial-to-mesenchymal transition (EMT) in BC. Here, we aim to explore the role of miR-671-5p in the progression of BC oncogenic transformation and treatment. METHODS: The 21T series cell lines, which were originally derived from the same patient with metastatic BC, including normal epithelia (H16N2), ADH (21PT), primary DCIS (21NT), and cells derived from pleural effusion of lung metastasis (21MT), and human BC specimens were used. Microdissection, miRNA transfection, dual-luciferase, radio- and chemosensitivity, and host-cell reactivation (HCR) assays were performed. RESULTS: Expression of miR-671-5p displays a gradual dynamic decrease from ADH, to DCIS, and to IDC. Interestingly, the decreased expression of miR-671-5p detected in ADH coexisted with advanced lesions, such as DCIS and/or IDC (cADH), but not in simple ADH (sADH). Ectopic transfection of miR-671-5p significantly inhibited cell proliferation in 21NT (DCIS) and 21MT (IDC), but not in H16N2 (normal) and 21PT (ADH) cell lines. At the same time, the effect exhibited in time- and dose-dependent manner. Interestingly, miR-671-5p significantly suppressed invasion in 21PT, 21NT, and 21MT cell lines. Furthermore, miR-671-5p suppressed FOXM1-mediated EMT in all 21T cell lines. In addition, miR-671-5p sensitizes these cell lines to UV and chemotherapeutic exposure by reducing the DNA repair capability. CONCLUSIONS: miR-671-5p displays a dynamic decrease expression during the oncogenic transition of BC by suppressing FOXM1-mediated EMT and DNA repair. Therefore, miR-671-5p may serve as a novel biomarker for early BC detection as well as a therapeutic target for BC management.

miR-638 mediated regulation of BRCA1 affects DNA repair and sensitivity to UV and cisplatin in triple-negative breast cancer.

INTRODUCTION: Triple-negative breast cancer (TNBC) represents 15 to 20% of all types of breast cancer; however, it accounts for a large number of metastatic cases and deaths, and there is still no effective treatment. The deregulation of microRNAs (miRNAs) in breast cancer has been widely reported. We previously identified that miR-638 was one of the most deregulated miRNAs in breast cancer progression. Bioinformatics analysis revealed that miR-638 directly targets BRCA1. The aim of this study was to investigate the role of miR-638 in breast cancer prognosis and treatment. METHODS: Formalin-fixed, paraffin-embedded (FFPE) breast cancer samples were microdissected into normal epithelial and invasive ductal carcinoma (IDC) cells, and total RNA was isolated. Several breast cancer cell lines were used for the functional analysis. miR-638 target genes were identified by TARGETSCAN-VERT 6.2 and miRanda. The expression of miR-638 and its target genes was analyzed by real-time qRT-PCR and Western blotting. Dual-luciferase reporter assay was employed to confirm the specificity of miR-638 target genes. The biological function of miR-638 was analyzed by MTT chemosensitivity, matrigel invasion and host cell reactivation assays. RESULTS: The expression of miR-638 was decreased in IDC tissue samples compared to their adjacent normal controls. The decreased miR-638 expression was more prevalent in non-TNBC compared with TNBC cases. miR-638 expression was significantly downregulated in breast cancer cell lines compared to the immortalized MCF-10A epithelial cells. BRCA1 was predicted as one of the direct targets of miR-638, which was subsequently confirmed by dual-luciferase reporter assay. Forced expression of miR-638 resulted in a significantly reduced proliferation rate as well as decreased invasive ability in TNBC cells. Furthermore, miR-638 overexpression increased sensitivity to DNA-damaging agents, ultraviolet (UV) and cisplatin, but not to 5-fluorouracil (5-FU) and epirubicin exposure in TNBC cells. Host cell reactivation assays showed that miR-638 reduced DNA repair capability in post UV/cisplatin-exposed TNBC cells. The reduced proliferation, invasive ability, and DNA repair capabilities are associated with downregulated BRCA1 expression. CONCLUSIONS: Our findings suggest that miR-638 plays an important role in TNBC progression via BRCA1 deregulation. Therefore, miR-638 might serve as a potential prognostic biomarker and therapeutic target for breast cancer.

Evaluation of Contralateral Limb Cross Education and High-Frequency Repetitive Transcranial Magnetic Stimulation on Functional Indices of the Affected Upper Limb in Subacute Phase of Stroke.

Background: Stroke is one of the causes of long-term morbidity. Despite rehabilitation strategies, most survivors live with motor deficits in the upper limbs. Objectives: The aim of the study was to compare the effect of contralateral cross education (CE) and high-frequency repetitive magnetic stimulation (HF-rTMS) on the function of upper extremity in subacute phase of stroke. Methods: Forty patients were randomly assigned into 4 groups. Group "A" received physical therapy (PT) for 10 sessions, 3 times per week. Group "B" received PT and HF-rTMS as follows: stimulation of 20 Hz for 5 s, intertrain interval for 50 s, 20 trains, 2000 pulses at 90% resting motor threshold, and conventional PT. Group "C" was treated with CE and PT. In group "D," HF-rTMS, CE, and PT were administered. Results: Significant differences were found in the Fugl-Meyer scale between "A" and "C" (P = 0.01), "A" and "D" (P = 0.02), and "B" and "C" groups (P = 0.01). In the box-block test, there were significant differences between "A" and "B" (P = 0.01), "A" and "C" (P < 0.001), "B" and "D" (P = 0.001), and "B" and "C" groups (P = 0.01). Statistical differences were observed in grip strength between "A" and "B" (P = 0.01) and "A" and "C" groups (P = 0.02). Conclusions: It is suggested that clinicians select the therapeutic methods in line with their expected goal. When the goal is to improve upper extremity function, CE+PT could be more effective than HF-rTMS+PT. Also, CE+PT and HF-rTMS+PT were more effective than CE+HF-rTMS+PT at improving grip strength. Therefore, combining several methods would not always lead to better results.

A Comparison between Effects of Neurofeedback and Balance Exercise on Balance of Healthy Older Adults.

BACKGROUND: Balance ability is a crucial component of independent daily activities among the older adultss. Balance impairment is one of the major risk factors for falls and related complications. OBJECTIVE: The present study aims to investigate and compare the effect of neurofeedback training and balance training on balance and fall risk among older adults. MATERIAL AND METHODS: In this randomized controlled trial, a total of 48 older adults aged more than 65 years were recruited and randomly assigned into two groups, neurofeedback group (n=24) and balance exercise group (n=24). Prior to the intervention, the static balance, dynamic balance, and fall risk were measured using Biodex D balance system and Fullerton Advanced Balance scale. Subjects in neurofeedback group received neurofeedback training for 12 sessions of 30-min, every other day. Moreover, subjects in balance exercise group received balance training for four weeks in 12 sessions (45-minute) every other day. After the intervention, balance measurements were repeated in both groups. The significance level was set at p<0.05. RESULTS: Static balance and dynamic balance were shown to significantly improve, after the interventions (p<0.001). Furthermore, fall risk was significantly reduced, after the trial (p<0.001). In addition, the therapeutic effect of neurofeedback training was not less significant than exercises on balance in the older adults (p<0.001). CONCLUSION: The findings suggest that both neurofeedback training and balance training improved balance ability among the older adults. Results also show the therapeutic effect of neurofeedback training on balance in older people. However, further research is required to accurately investigate the long-term effects of these two treatment methods among the older adults.

Use of the T-1470 LiteTouch™ Laser in the En Bloc Resection of an Upper Tract Urothelial Cancer.

BACKGROUND: Endoscopic laser-ablative therapy of upper tract urothelial carcinoma offers kidney-sparing treatment for well-selected low-risk tumors. The traditional technique consists of tumor biopsy with flexible forceps or nitinol basket for pathologic assessment of stage and grade, followed by laser ablation of the tumor. In this case, we present the use of the new T-1470 LiteTouch™ laser for intraoperative tumor en bloc resection, affording both tissue acquisition and tumor ablation. Case Presentation. An 81-year-old female with a past medical history significant for stage 4 chronic kidney disease, peripheral artery disease, coronary artery disease, type 2 diabetes mellitus, and gout was diagnosed with a 2 cm left upper tract high-grade papillary urothelial carcinoma confirmed by cytology with cell block preparation. Using a novel approach, the tumor was resected, en bloc, using the T-1470 LiteTouch™ laser which allowed for sufficient tissue resection for pathologic examination and strong hemostasis. This new technique is the first recorded example of tumor en bloc resection using the T-1470 LiteTouch™ laser of an upper tract urothelial carcinoma. CONCLUSION: The use of the T-1470 LiteTouch™ laser offers promise for its use as a novel laser for the endoscopic treatment of upper tract urothelial carcinoma. It shows potential for advantages over current techniques through its ability to achieve en bloc resection and superior hemostasis.

The Effect of High-Frequency Repetitive Transcranial Magnetic Stimulation on Functional Indices of Affected Upper Limb in Patients with Subacute Stroke.

BACKGROUND: Repetitive transcranial magnetic stimulation (rTMS) is a novel technique that may improve recovery in patients with stoke, but the role of rTMS as an applied and practical treatment modality for stroke rehabilitation has not been established yet. OBJECTIVE: This study was conducted to determine the effects of a rehabilitation program (RP) in conjunction with rTMS on functional indices of the paretic upper limb in the subacute phase of stroke. MATERIAL AND METHODS: In this experimental study, twenty patients in the subacute phase of stroke were randomly assigned into two groups: The high frequency rTMS (HF-rTMS) in conjunction with RP (experimental group), and the RP group (control group). The experimental group received 10 sessions of 20 Hz rTMS on the affected primary motor cortex and the other group received 10 sessions of RP. In experimental group, RP for the paretic hand was conducted following rTMS session. Box and block test (BBT), Fugl-Meyer Motor Assessment for upper limb (FMA-UL), grip strength and pinch strength were used to assess motor function before the first session and after the last session of treatment. RESULTS: Significant improvement in BBT, FMA-UL, grip strength and pinch strength was observed in both groups. Improvement of BBT and grip strength was significantly greater in the experimental group rather than the control group (p<0.05). FMA-UL score and the pinch strength were greater in the experimental group, although the differences were not statistically significant. CONCLUSION: HF-rTMS in conjunction with RP is effective to improve the function of upper limb. It seems HF-rTMS is a novel feasible and safe technique for hemiparesis patients in the subacute phase of stroke.

Tumor immune microenvironment in non-muscle-invasive urothelial carcinoma of the bladder.

Immunotherapy has gained significance in a variety of tumor types including advanced urothelial carcinoma. Noninvasive urothelial lesions have been treated with intravesical Bacillus-Calmette-Guerin (BCG) for decades. Given treatment failure in a subset of these tumors, ongoing clinical trials investigating the role of checkpoint inhibitors are actively pursued in this group of patients. The present study aims to delineate PD-L1, CD8, and FOXP3 expression in tumor microenvironment in non-muscle-invasive urothelial carcinoma samples obtained via sequential biopsies and to assess its potential role in predicting disease outcome. Cases with >1% and> 5% PD-L1 expression in tumor cells showed lower relative risk (RR) to recur at any subsequent biopsy compared with those with lower PD-L1 expression (RRs, 0.83 [P = .009] and 0.81 [P = .03], respectively). Cases with higher expression of FOXP3 in peritumoral lymphocytes were at lower risk for tumor grade progression at any biopsy (RR, 0.2; P = .02). Tumors with FOXP3/CD8 expression ratio of >1 in intratumoral lymphocytes had lower risk of grade progression (RR, 0.28; P = .04). Although higher number of FOXP3-, CD8-, and PD-L1-positive lymphocytes were encountered after BCG treatment, the findings did not reach statistical significance. In patients without BCG treatment, PD-L1 expression in tumor cells and peritumoral lymphocytes varied across serial biopsies, suggesting the need for additional approaches to assess eligibility for immunotherapy in non-muscle-invasive urothelial carcinoma patients.

A Role for De Novo Purine Metabolic Enzyme PAICS in Bladder Cancer Progression.

Genomic and transcriptome sequencing of bladder cancer (BLCA) has identified multiple molecular alterations during cancer progression. Many of these identified genetic and epigenetic changes play a role in the progression of this disease. Studies have identified molecular subtypes in muscle-invasive bladder cancer (MIBC) with different sensitivities to frontline therapy suggesting the heterogeneity in these tumors and the importance of molecular characterization of MIBC to provide effective treatment. Specifically, it has become increasingly evident, as demonstrated by The Cancer Genome Atlas project, that metabolic enzymes are commonly dysregulated in BLCA. Elevated expression of multiple metabolic enzymes is due to the increased demand from rapidly proliferating BLCA cells requiring extensive nucleotide synthesis. Cancer cells utilize the de novo purine and pyrimidine biosynthetic pathway as a source of their nucleotide needs. In this study, we show that phosphoribosyl aminoimidazole succinocarboxamide synthetase (PAICS), an enzyme involved in de novo purine biosynthetic pathway, is significantly overexpressed in BLCA. Immunohistochemical staining of paraffin-embedded tissue sections showed that PAICS is overexpressed in MIBC. Furthermore, we found that tumor suppressor miR-128 negatively regulated PAICS expression by binding to its 3'-untranslated region. We also found that PAICS induces EMT by positively regulating SNAI1 and by a reduction in E-cadherin expression. Additionally, our in vitro functional studies and in vivo chicken chorioallantoic membrane assay show that PAICS plays a critical role in BLCA cell proliferation, invasion, and tumor growth. Collectively, our data suggest that targeting PAICS may provide a therapeutic option in BLCA.

Choriocarcinoma with brain, lung and vaginal metastases successfully treated without brain radiation or intrathecal chemotherapy: A case report.

•There is no consensus on optimal treatment for GTN and brain metastases.•Brain metastasis treated with craniotomy and intravenous, EMA-CO chemotherapy•Intravenous high-dose methotrexate may be adequate to treat brain metastases.

miR-671-5p inhibits epithelial-to-mesenchymal transition by downregulating FOXM1 expression in breast cancer.

MicroRNA (miRNA) dysfunction is associated with a variety of human diseases, including cancer. Our previous study showed that miR-671-5p was deregulated throughout breast cancer progression. Here, we report for the first time that miR-671-5p is a tumor-suppressor miRNA in breast tumorigenesis. We found that expression of miR-671-5p was decreased significantly in invasive ductal carcinoma (IDC) compared to normal in microdissected formalin-fixed, paraffin-embedded (FFPE) tissues. Forkhead Box M1 (FOXM1), an oncogenic transcription factor, was predicted as one of the direct targets of miR-671-5p, which was subsequently confirmed by luciferase assays. Forced expression of miR-671-5p in breast cancer cell lines downregulated FOXM1 expression, and attenuated the proliferation and invasion in breast cancer cell lines. Notably, overexpression of miR-671-5p resulted in a shift from epithelial-to-mesenchymal transition (EMT) to mesenchymal-to-epithelial transition (MET) phenotypes in MDA-MB-231 breast cancer cells and induced S-phase arrest. Moreover, miR-671-5p sensitized breast cancer cells to cisplatin, 5-fluorouracil (5-FU) and epirubicin exposure. Host cell reactivation (HCR) assays showed that miR-671-5p reduces DNA repair capability in post-drug exposed breast cancer cells. cDNA microarray data revealed that differentially expressed genes when miR-671-5p was transfected are associated with cell proliferation, invasion, cell cycle, and EMT. These data indicate that miR-671-5p functions as a tumor suppressor miRNA in breast cancer by directly targeting FOXM1. Hence, miR-671-5p may serve as a novel therapeutic target for breast cancer management.

Utility of uroplakin II expression as a marker of urothelial carcinoma.

Uroplakins are markers of terminally differentiated urothelium. Uroplakin II (UPII) is a newly described sensitive marker for urothelial carcinoma (UC). The expression profile of UPII in different types of UC and its utility in the diagnostic setting are needed. We evaluated UPII expression in bladder tissue microarrays, including urothelial neoplasm of low malignant potential (n = 8), low-grade papillary UC (n = 72), noninvasive high-grade papillary UC (n = 77), UC in situ (n = 27), and invasive high-grade UC (INVUC) (n = 122). UPII expression in 52 breast carcinomas and 38 high-grade prostate adenocarcinomas was also assessed. UPII expression was compared with GATA binding protein 3 (GATA3) and estrogen receptor for its role in facilitating the differential diagnosis of the above 3 types of malignancy. UPII labeling was seen in 83.0% of UC overall, including 95.7% of noninvasive UC and 65.6% of INVUC. UPII labeling was not found in any breast and prostate carcinomas. In comparison, GATA3 labeling was seen in 91.6% of all UCs, including 96.4% of noninvasive UCs and 85.1% of INVUC, with stronger intensity and extent compared with UPII (P < .005). GATA3 labeled 2 (5%) of 38 high-grade prostate adenocarcinoma. Estrogen receptor nuclear labeling was seen in 13.0% of UCs and 12.5% of prostate carcinomas. UPII was highly specific (100%) but only moderately sensitive for UC and can therefore be a potentially useful marker to identify urothelial lineage and help distinguish UC from prostate cancer or, in conjunction with GATA3, from metastatic breast cancer.

MicroRNA-21 Down-regulates Rb1 Expression by Targeting PDCD4 in Retinoblastoma.

Retinoblastoma (RB) is a children's ocular cancer caused by mutated retinoblastoma 1 (Rb1) gene on both alleles. Rb1 and other related genes could be regulated by microRNAs (miRNA) via complementarily pairing with their target sites. MicroRNA-21 (miR-21) possesses the oncogenic potential to target several tumor suppressor genes, including PDCD4, and regulates tumor progression and metastasis. However, the mechanism of how miR-21 regulates PDCD4 is poorly understood in RB. We investigated the expression of miRNAs in RB cell lines and identified that miR-21 is one of the most deregulated miRNAs in RB. Using qRT-PCR, we verified the expression level of several miRNAs identified by independent microarray assays, and analyzed miRNA expression patterns in three RB cell lines, including Weri-Rb1, Y79 and RB355. We found that miR-19b, -21, -26a, -195 and -222 were highly expressed in all three cell lines, suggesting their potential role in RB tumorigenesis. Using the TargetScan program, we identified a list of potential target genes of these miRNAs, of which PDCD4 is one the targets of miR-21. In this study, we focused on the regulatory mechanism of miR-21 on PDCD4 in RB. We demonstrated an inverse correlation between miR-21 and PDCD4 expression in Weri-Rb1 and Y79 cells. These data suggest that miR-21 down-regulates Rb1 by targeting PDCD4 tumor suppressor. Therefore, miR-21 could serve as a therapeutic target for retinoblastoma.

Serum glutamate levels correlate with Gleason score and glutamate blockade decreases proliferation, migration, and invasion and induces apoptosis in prostate cancer cells.

PURPOSE: During glutaminolysis, glutamine is catabolized to glutamate and incorporated into citric acid cycle and lipogenesis. Serum glutamate levels were measured in patients with primary prostate cancer or metastatic castrate-resistant prostate cancer (mCRPCa) to establish clinical relevance. The effect of glutamate deprivation or blockade by metabotropic glutamate receptor 1 (GRM1) antagonists was investigated on prostate cancer cells' growth, migration, and invasion to establish biologic relevance. EXPERIMENTAL DESIGN: Serum glutamate levels were measured in normal men (n = 60) and patients with primary prostate cancer (n = 197) or mCRPCa (n = 109). GRM1 expression in prostatic tissues was examined using immunohistochemistry (IHC). Cell growth, migration, and invasion were determined using cell cytotoxicity and modified Boyden chamber assays, respectively. Apoptosis was detected using immunoblotting against cleaved caspases, PARP, and γ-H2AX. RESULTS: Univariate and multivariate analyses showed significantly higher serum glutamate levels in Gleason score ≥ 8 than in the Gleason score ≤ 7 and in African Americans than in the Caucasian Americans. African Americans with mCRPCa had significantly higher serum glutamate levels than those with primary prostate cancer or benign prostate. However, in Caucasian Americans, serum glutamate levels were similar in normal research subjects and patients with mCRPC. IHC showed weak or no expression of GRM1 in luminal acinar epithelial cells of normal or hyperplastic glands but high expression in primary or metastatic prostate cancer tissues. Glutamate deprivation or blockade decreased prostate cancer cells' proliferation, migration, and invasion and led to apoptotic cell death. CONCLUSIONS: Glutamate expression is mechanistically associated with and may provide a biomarker of prostate cancer aggressiveness.