LncRNA SNHG8 promotes cell migration and invasion in breast cancer cell through miR-634/ZBTB20 axis.

OBJECTIVE: Small nucleolus RNA Host Gene 8 (SNHG8) belongs to a subgroup of long non-coding RNAs. SNHG8 is upregulated in many cancers, such as gastric cancer, liver cancer, and esophageal squamous cell cancer. However, whether SNHG8 is abnormally expressed in breast cancer and its biological functions remain unclear. Therefore, our research intended to determine the expression status of SNHG8 in breast cancer, explore the effects of SNHG8 on the development of breast cancer, and investigate the potential molecular mechanisms in cancer progression. PATIENTS AND METHODS: The expression levels of SNHG8 were detected in tissue samples and cell lines via qRT-PCR. The effects of SNHG8 on viability of breast cancer cells were detected via CCK-8, EdU, transwell, and flow cytometry analyses. RESULTS: qRT-PCR results showed that the expression level of SNHG8 was significantly upregulated in tumor tissues and cell lines. Gene functional studies showed that the downregulation of the expression level of SNHG8 significantly inhibited the breast cancer cells migration and invasion, and induced apoptosis. Meanwhile, we found that SNHG8 served as an inhibitor of miR-634 in tumor tissues. SNHG8 may participate in the malignancy of breast cancer by sponging the miR-634 to increase the expression level of ZBTB20. CONCLUSIONS: The SNHG8-miR-634-ZBTB20 pathway may be a potential target for the treatment of breast cancers.

[Tap-hammer elicited vestibular-evoked myogenic potentials system: its design and preliminary application].

Objective: The aim of this study was to design and perform "Tap-hammer"system that can be used to elicit vestibular evoked myogenic potentials (VEMP) in normal adults and to report the preliminary results of this system. Methods: A triggered Tap-hammer was designed, made and connected with an electric recording system, to form as a system for Tap-VEMP recording. Twenty healthy adult volunteers (7 males and 13 females, aged 20 to 37 years, 40 ears in total) were recruited for air-conducted sound VEMP (ACS-VEMP) and Tap-VEMP examinations. Waveforms and parameters of both VEMPs were recorded and analyzed. SPSS 22.0 software was used for statistical analysis. Results: The response rates of ACS-, Tap-ocular VEMP (oVEMP) and ACS-, Tap-cervical VEMP (cVEMP) were both 100% (40/40). The mean±SD n1 latency, p1 latency, n1-p1 interval, amplitude, and asymmetry ratio (AR%) of Tap-oVEMP were (9.80±2.51)ms, (13.90±3.26)ms, (4.09±1.43)ms, (16.43±9.61)µV, (22.68±17.35)% respectively. The mean±SD p1 latency, n1 latency, p1-n1 interval, amplitude, and asymmetry ratio (AR%) of Tap-cVEMP were (13.26±2.07)ms, (21.84±2.89)ms, (8.58±2.10)ms, (457.65±274.94)µV, (20.42±13.46)% respectively. Both n1 latency and p1 latency of Tap-VEMPs were shorter than those in ACS-VEMPs (P<0.05). No statistical difference could be found between the two stimulation methods in the parameters of n1-p1 interval, amplitude, and asymmetry ratio(P>0.05). Conclusion: The Tap-hammer system can elicit VEMP responses in healthy young people. This system can be used as an alternative stimulation method for bone conduction VEMP.

Circ_001680 aggravates the malignant process of gastric carcinoma by targeting MAP2.

OBJECTIVE: This study aims to explore the expression pattern and clinical significance of circ_001680 in gastric carcinoma (GC) process. PATIENTS AND METHODS: Circ_001680 levels in 40 pairs of GC and paracancerous ones were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The relationship between circ_001680 and GC clinicopathological parameters was analyzed. AGS and SGC-7901 cells were used for constructing circ_001680 knockdown models by shRNA transfection. Proliferative and metastatic abilities in GC cells with circ_001680 knockdown were examined by cell counting kit-8 (CCK-8) and transwell assay, respectively. Dual-Luciferase reporter assay was conducted to clarify the interaction between circ_001680 and MAP2. Their co-regulation on GC process was detected through rescue experiments. RESULTS: Circ_001680 was highly expressed in GC tissues and cell lines. High level of circ_001680 predicted high incidences of lymphatic and distant metastasis, and poor prognosis in GC patients. Knockdown of circ_001680 suppressed proliferative and metastatic abilities in AGS and SGC-7901 cells. MAP2 was the target gene binding circ_001680, which was lowly expressed in GC. In addition, MAP2 was negatively correlated to circ_001680. Knockdown of MAP2 could abolish the suppressed proliferative and metastatic abilities in GC cells with circ_001680 knockdown. CONCLUSIONS: Circ_001680 is highly expressed in GC tissues and closely related to metastasis and prognosis in GC patients, which promotes the proliferative and metastatic abilities in GC cells by negatively interacting with MAP2.

Overall adjustment acupuncture for postmenopausal osteoporosis (PMOP): a study protocol for a randomized sham-controlled trial.

BACKGROUND: Osteoporosis is becoming more prevalent in aging societies worldwide, and the economic burden attributable to osteoporotic fractures is substantial. The medications presently available to treat osteoporosis have side effects. Acupuncture is widely used for treating osteoporotic postmenopausal women because it is non-invasive and has fewer side effects, but the powerful clinical evidence for its efficacy remains insufficient. Our study intends to explore the effect of overall adjustment acupuncture (OA) in the treatment of postmenopausal osteoporosis (PMOP). METHODS/DESIGN: This study is a randomized, sham-controlled, patient- and assessor-blinded trial and aims to evaluate the effect of OA in women with PMOP. We will recruit 104 women aged 45-70 years with a diagnosis of PMOP. Participants will be randomly allocated in a 1:1 ratio to the OA group and the sham acupuncture (SA) group. Both groups will receive real herbal medicine treatment as a basic treatment twice a day for 3 months, the OA group receives real acupuncture treatment and the SA group receives placebo acupuncture treatment (non-penetrating, sham skin-needle therapy, sham cupping). All patients will receive acupuncture treatment twice per week for 3 months. The primary outcome is bone mineral density (BMD) and the secondary outcomes include estradiol (E2), follicle-stimulating hormone (FSH), bone gla protein (BGP), bone alkaline phosphatase (BALP), total antioxidant capacity (TAC), advanced oxidation protein products (AOPP), PPARγ, ß-catenin, FoxO3a levels, visual analog pain scale score (VAS), Traditional Chinese medicine (TCM) syndrome scores and quality of daily life score (QOL). Outcome measures will be collected at baseline, middle of the treatment (1.5 months), the end of treatment (3 months). The present protocol followed the SPIRIT guidelines and fulfills the SPIRIT Checklist. CONCLUSION: This study will be conducted to compare the efficacy of OA versus SA. This trial should help to evaluate whether OA can effectively prevent and treat PMOP by improving the estrogen levels of postmenopausal women. The mechanism is to improve the imbalance of osteogenic differentiation and lipogenesis of bone-marrow cells under oxidative stress. TRIAL REGISTRATION: Chinese Clinical Trial Registry, ID: ChiCTR1800017581. Registered on 5 August 2018. URL: http://www.chictr.org.cn.

MiR-593-5p promotes the development of hypoxic-induced pulmonary hypertension via targeting PLK1.

OBJECTIVE: The aim of this study was to investigate the role of microRNA-593-5p (miR-593-5p) in hypoxia-induced pulmonary hypertension (HPH), and to explore its underlying mechanism. MATERIALS AND METHODS: Sprague-Dawley (SD) rats were housed in a hypoxia environment 8 hours per day for consecutive 4 weeks. After the establishment of the HPH rat model, we detected the right ventricular systolic pressure (RVSP) and right heart hypertrophy index (RVHI) in HPH rats and controls. The expression levels of miR-593-5p and polo-like kinase 1 (PLK1) in rat lungs were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Subsequently, miR-593-5p mimics and inhibitor were constructed and transfected into cells. The proliferation and migration of pulmonary arterial smooth muscle cells (PASMCs) were accessed by cell counting kit-8 (CCK-8) assay and wound healing assay, respectively. The protein level of PLK1 in PASMCs after transfection with miR-593-5p mimics or inhibitor was detected by Western blot. Dual-luciferase reporter gene assay was conducted to verify the binding condition of miR-593-5p and PLK1. Finally, rescue experiments were performed to explore whether the regulatory effect of miR-593-5p on HPH development was associated with PLK1. RESULTS: RVSP and RVHI in rats of the hypoxic group were significantly higher than those of controls. MiR-593-5p was significantly downregulated while PLK1 was remarkably upregulated in lung tissues of HPH rats than those of controls. Similarly, miR-593-5p expression in PASMCs decreased gradually with the prolongation of hypoxia induction. Overexpression of miR-593-5p remarkably inhibited the proliferation and migration of PASMCs. Subsequently, dual-luciferase reporter gene verified the binding condition of miR-593-5p and PLK1. Both the mRNA and protein levels of PLK1 were negatively regulated by miR-593-5p. Also, rescue experiments demonstrated that the inhibitory effects of miR-593-5p on the proliferation and migration of PASMCs could be reversed by PLK1 overexpression. CONCLUSIONS: MiR-593-5p is lowly expressed in lung tissues of HPH rats. Meanwhile, it stimulates the proliferation and migration of PASMCs via targeting PLK1, thereby promoting HPH development.

CD151 promotes breast cancer metastasis by activating TGF-ß1/Smad signaling pathway.

OBJECTIVE: This study aimed to explore the expression characteristics of CD151 in breast cancer (BC) and to further study its role in the development of BC and potential regulatory mechanisms. PATIENTS AND METHODS: Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) was used to detect the level of CD151 in 82 pairs of BC tissues and adjacent normal ones, and the relationship between CD151 expression and BC pathological parameters and prognosis was analyzed. CD151 expression in BC cells was further validated using qRT-PCR. The CD151 knockdown model was constructed in BC cell lines including MCF-7 and SKBR3 using the small interference RNA. The cell counting kit-8 (CCK-8) and transwell assay were used to analyze the effect of CD151 on the biological function of BC cells, and finally Western blot was performed to explore its underlying mechanism. RESULTS: QRT-PCR analysis revealed that CD151 level in BC tissues was strikingly higher than that in normal ones, and the difference was statistically significant. Compared with patients with low CD151 level, patients with high CD151 level had worse tumor stage, lymph node metastasis, and distant metastases. The higher the incidence of metastasis, the lower the overall survival rate. Compared with the negative control group, the ability of cell proliferation or invasion and migration in the CD151 knockdown group was significantly reduced. In addition, Western blot results demonstrated that the levels of proteins in TGF-ß1/Smad pathway, including transforming growth factor-ß1 (TGF-ß1), p-Smad2, p-Smad3, N-cad, Vimentin and MMP-9, were remarkably decreased in cells of si-CD151 group. CONCLUSIONS: The expression of CD151 in BC was significantly increased, which was found evidently associated with BC stage, lymph node or distant metastasis, and poor prognosis. Meanwhile, CD151 may promote the proliferation and invasion of BC by regulating TGF-ß1/Smad pathway.

Expression of mTOR and its inhibitory effect on cell proliferation and apoptosis of breast cancer cells.

The aim of this study is to investigate the expression of mTOR in breast cancer and observe the effect of CCI-779 on proliferation and apoptosis of MDA-MB-231 cells. Immunohistochemical staining was used to detect the expression of mTOR protein in breast cancer tissues and MDA-MB-231 cells. MTT assay was used to assess the effect of CCI-779 on proliferation of MDA-MB-231 cells. Annex-inV-FITC/ PI assay was utilized to evaluate the effect of CCI-779 on apoptosis of MDA-MB-231 cells. Among the 71 cases of breast cancer tissues, 54.9% were mTOR-positive that exhibited significantly higher expression than the 32 cases of normal tissues (21.9%); mTOR protein was also found to be expressed in MDA-MB-231 cells. The mTOR inhibitor CCI-779 significantly inhibited the proliferation of MDA-MB-231 cells that was dose- and time-dependent. However, CCI-779 was unable to induce apoptosis of MDA-MB-231 cells as demonstrated with AnnexinV-FITC/PI assay. mTOR plays a key role in the initiation and development of breast cancer, and its inhibitor CCI-779 exerts a strong suppressive activity against MDA-MB-231 cells, suggesting its therapeutic potential to treat breast cancer.

The role of Sep (O-phosphoserine) tRNA: Sec (selenocysteine) synthase (SEPSECS) in proliferation, apoptosis and hormone secretion of trophoblast cells.

OBJECTIVES: To investigate whether Sep (O-phosphoserine) tRNA: Sec (selenocysteine) synthase (SEPSECS), which plays an essential role in the synthesis of selenoprotein, affects proliferation, apoptosis and hormone secretion of human trophoblast cells. METHODS: Human trophoblast JEG-3 cells were divided into four groups: control group, SEPSECS silenced-expression group, empty vector group and SEPSECS over-expression group. Over-expression and silenced-expression were achieved by transfection with plasmid DNA or RNA oligonucleotide, respectively. 3-[4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide (MTT) and colony formation assays were performed to investigate cell proliferation, while apoptosis was tested by annexin V-FITC, PI double staining and caspases-3 activation assays, enzyme-linked immunosorbent assay (ELISA) was used to determine the level of progesterone (PG) and human chorionic gonadotropin (hCG). RESULTS: SEPSECS silenced-expression clearly inhibited proliferation of JEG-3 cells (p < 0.05), significantly induced cell apoptosis (p < 0.01) and reduced the production of PG and hCG (p < 0.05). On the contrary, SEPSECS over-expression significantly promoted both cell proliferation (p < 0.01) and secretion of PG and hCG (p < 0.05). CONCLUSIONS: SEPSECS significantly affects proliferation, apoptosis and hormone secretion of human trophoblast cells, suggesting that a potential relationship exists among SEPSECS, cell proliferation, apoptosis and hormone production of human placental trophoblast cells. Furthermore, this may provide a clue to uncover the relationship between selenium and human placental in association with an emphasis on the importance of selenium adequacy during pregnancy.