MiR-506 suppresses papillary thyroid carcinoma cell proliferation and metastasis via targeting IL17RD.

OBJECTIVE: The aim of this study was to determine the role of microRNA-506-3p (miR-506) in papillary thyroid carcinoma (PTC), and to further explore the underlying mechanism. PATIENTS AND METHODS: The expression level of miR-506 in clinical cases was detected by Real Time-fluorescence quantitative Polymerase Chain Reaction (RT-qPCR). Meanwhile, RT-qPCR was performed to determine miR-506 expression in different PTC cell lines. Bioinformatics software was used to predict the possible target genes of miR-506. Dual-Luciferase reporter gene assay together with Western blot (WB) assay were used to verify the prediction results. Finally, cellular functions such as proliferation and metastasis capacities were detected in vitro. RESULTS: RT-qPCR was used to measure the expression level of miR-506 in 80 paired PTC cases. The results showed that the expression level of miR-506 in PTC tissues was significantly decreased. In vitro, miR-506 expression was also markedly suppressed in four PTC cell lines. TPC-1 cells expressed the lowest level of miR-506. Subsequently, the target gene of miR-506 was predicted by TargetScan, miRBase and miRanda. The prediction results indicated that IL17RD was an alternative target gene of miR-506. Furthermore, miR-506 was found to remarkably inhibit the Luciferase activity of wild-type IL17RD. However, it had no effect on mutant-type. Besides, the protein expression level of IL17RD was significantly reduced in miR-506-overexpressing TPC-1 cells. More importantly, the restored expression of IL17RD could alleviate the blocking effects of miR-506 on cell proliferation, migration and invasion. CONCLUSIONS: In this study, we found that miR-506 could inhibit the proliferation and metastasis of PTC cells. Meanwhile, IL17RD might be a downstream target of the biological process. Our findings provided a new therapeutic direction for the treatment of PTC.

MiR-155 promotes the proliferation and migration of breast cancer cells via targeting SOCS1 and MMP16.

OBJECTIVE: The aim of this study was to investigate the effect of miR-155 on the proliferation and migration of breast cancer cells, and to explore the underlying mechanism. MATERIALS AND METHODS: The breast cancer cell line MDA-MB-231 was transfected with miR-155 mimics, inhibitor or negative control, respectively. The expression level of miR-155 was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Subsequently, the proliferation of MDA-MB-231 cells was detected by multi-cellular tumor spheroid (MTS) and colony formation assay. Cell migration was examined by transwell assay and scratch test. In addition, qRT-PCR was performed to analyze the expression of matrix metallopeptidase 16 (MMP16) after miR-155 mimics or inhibitor transfection in MDA-MB-231 cells. Meanwhile, Western blot was used to evaluate the protein expression levels of suppressor of cytokine signaling 1 (SOCS1) and MMP16 after miR-155 mimics or inhibitor transfection. RESULTS: QRT-PCR results showed that miR-155 mimics significantly increased the expression of miR-155 in MDA-MB-231 cells, whereas miR-155 inhibitor markedly decreased miR-155 expression (p < 0.05). Meanwhile, MTS and colony formation assay indicated that the proliferation of MDA-MB-231 cells was remarkably increased after miR-155 mimics transfection. However, miR-155 inhibitor transfection exhibited the opposite result in cell proliferation (p < 0.05). Moreover, miR-155 overexpression significantly increased the migration of MDA-MB-231 cells (p < 0.05). Western blot further confirmed that miR-155 overexpression down-regulated the expression level of target protein SOCS1 and upregulated the expression level of MMP16. CONCLUSIONS: We found that miR-155 significantly enhanced the proliferation and migration of MDA-MB-231 cells, which might serve as an oncogene in breast cancer. Therefore, it is preliminarily believed that miR-155 plays an important role in the proliferation and migration of breast cancer cells via down-regulating the expression of SOCS1 and up-regulating the expression of MMP16.

IMP3 promotes TNBC stem cell property through miRNA-34a regulation.

OBJECTIVE: To explore the expression and function of insulin-like growth factor II (IGFII) mRNA binding protein (IMP3) in the Triple Negative Breast Cancer (TNBC). MATERIALS AND METHODS: According to previously reported gene expression array, we found that IMP3 had significantly higher expression in the CD44+CD24-ESA+ cell cluster, tumor initiating cell or cancer stem cell (CSCs), compared to other tumor cells. Based on the GEO database (GEO accession No. GSE6883), we detected the mRNA levels of IMP 1,2 and 3 by quantitative polymerase chain reaction (q-PCR) in CD44+CD24-ESA+ cell cluster and other breast tumor cell clusters. Besides, we measured IMP3 expression in microsphere of breast cancer, which exerted more significant tumor stem cell properties. The effects of IMP3 on breast cancer cell stem cell properties were studied by RNA interference and overexpression approaches in vitro. Furthermore, we predicted and identified microRNA, which could target and regulate IMP3 from bioinformatics analysis, and verified the interaction by luciferase assays and rescue experiments. RESULTS: Previously reported data showed that IMP3 expression was significantly upregulated in CD44+CD24-ESA+ cell cluster from breast cancer tissues. Besides, we found IMP3 had higher expression in mesenchymal cells rather than epithelial cells, which was also significantly elevated in SUM159 and T49D cell lines cultured as microsphere rather than adherent cells or differentiated cells. CD44+CD24-ESA+ cell cluster proportion was significantly decreased after silencing IMP3 in SUM1315, and its ability to develop into microsphere was significantly inhibited. By re-expressing IMP3 in SUM315, we restored the self-renewal capacity and tumorigenesis potential of SUM315. Through relative predicting website, we found several miRNAs which could regulate IMP3. miR-34a with highest score was chosen for further analysis. Mimicking miR-34a significantly downregulated IMP3 expression and inhibited its ability to develop into microsphere, while overexpressing IMP3 could rescue this process. CONCLUSIONS: IMP3 plays a vital role in maintaining stem cell properties of breast cancer cells, which could be regulated by mir-34a.

[Lack of progesterone receptor expression predicts poor prognosis in patients with operable ER-positive invasive breast cancer].

OBJECTIVE: To investigate the impact of lack of progesterone receptor (PR) expression on the prognosis of patients with operable ER (estrogen receptor)-positive invasive breast cancer. METHODS: We retrospectively analyzed the clinicopathological features, treatment and survival data of 318 women with ER+ /PR+ and ER+ /PR- invasive breast cancer. RESULTS: Among the 318 patients, there were 219 PR-positive and 99 PR-negative cases. The 5-year overall survival (OS) rate was 92.5%, and the 5-year disease-free survival (DFS) rate was 87.2% in the 318 ER-positive patients. Among them, the 5-year OS rates were significantly different between the PR-positive group (94.6%) and PR-negative group (87.8%, P=0.020), and the 5-year DFS rates were also significantly different from each other (89.8% and 81.6%, respectively, P=0.019). Univariate analysis showed that PR status, tumor size, T stage, axillary lymph node metastasis, and clinical stage were prognostic factors for OS (P<0.05 for all). Multivariate analysis showed that lack of PR expression, T stage ≥2, and positive axillary lymph node metastasis were independent risk factors for poor DFS and OS in ER-positive breast cancer patients (P<0.05 for all). Subgroup analysis showed that lack of PR expression was not significant in predicting poor DFS or OS when patients were in stage Ⅰ or with a small tumor (≤2 cm) (P>0.05 for all), and also showed that premenopausal women with PR-negative disease had poorer DFS and OS than PR-positive patients (P<0.05 for both). CONCLUSIONS: Lack of PR expression is an independent risk factor for poor prognosis in patients with operable ER-positive invasive breast cancer, especially in patients with a large tumor (>2 cm), advanced clinical stage (Stage Ⅱ or Ⅲ) or in premenopausal status.

The effective method for investigation meridian tropism theory in rats.

This present work describes an effective new method for study traditional Chinese medicine (TCM) on meridian tropism (MT) theory, which plays an essential role in clinical selection of TCM according to syndromes and strengthens the therapeutic effects. The new thread included material basis foundation and its tissue distribution study. Xiheliu, the most popular TCM on heart tropism, was investigated by simple and accurate high performance liquid chromatography (HPLC) method. The analysis of plasma after oral administration the total flavonoid of Xiheliu (TFX) exhibited that tamarixetin and kaempferide had the highest concentration and approximately the highest level within 25 min. The mixture of them could last accelerating the urine excretion more than 7 h after a single dose and could not cause the disorder of ion in rats, which was observed in diuretic activity experiment. In view of the reported biological activities was consistent with the effects of Xiheliu, tamarixetin and kaempferide were likely to be the material basis of it. Tissue distribution study showed that the highest level of analytes was in heart, lung, kidney and liver, and most tissues reached maximum level at 30 min post-dose. Since liver was the most important blood-supply tissue, the result of this experiment was in accordance with the MT record of Xiheliu and confirmed that tamarixetin and kaempferide was the material bases of it on MT. This is the first report for the illumination of material basis and the mechanism of Xiheliu on MT by analysis the record of Xiheliu in Compendium of Materia Medica and experimental study.

Diagnosis value of hysteroscopy for chronic endometritis.

OBJECTIVE: To investigate the correlation of hysteroscopy in the diagnosis of chronic endometritis (CE) with histology and assess its reliability. MATERIALS AND METHODS: Two hundred eleven patients with CE diagnosed by hysteroscopy were selected as the case group, and 30 cases without endometritis diagnosed by hysteroscopy were selected as the control group. Hysteroscopy and endometrial biopsy were carried out in all patients with endometrial hyperplasia. RESULTS: Among 211 patients with CE diagnosed by hysteroscopy, 200 cases were confirmed histologically. There was a significant correlation (p < 0.001) between characterization of CE by hysteroscopy and pathological grading. In 173 cases (86.5%), both histological and hysteroscopic grading were consistent (Kappa value = 0.62). CONCLUSION: Hysteroscopy is reliable in diagnosing CE and it can assess clinical effectiveness of antibiotic therapy.

Construction of three different recombinant scorpion fusion proteins with bifunctional activity.

This is the first report of three different fusion proteins with an antitumor-analgesic peptide obtained from Chinese scorpion Buthus martensii Karsch (BmKAGAP). The fusion proteins were constructed in the form of chimeric toxins, aiming to obtain bifunctional analgesic and antitumor activity. The fusion proteins consisted of luteinizing hormone-releasing hormone (LHRH), three different types of flexible linkers (L1, Ser-Ser-His-His-His-His-His-His-Ser-Ser-Gly-Leu-Val-Pro-Arg-Gly-Ser-His-Met; L2, Gly-Gly-Gly-Ser-Gly-Gly-Gly-Ser; L3, Ser-Gly-Gly-Ser-Gly-Gly-Ser-Gly-Gly-Gly-Ser-Ser-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser), and BmKAGAP. The genes coding three fusion proteins were cloned and expressed in E. coli in soluble form. Following two successive column chromatographic separations, purified fusion proteins were obtained. These fusion proteins exhibited analgesic activity in mice and were cytotoxic to a hepatocellular carcinoma cell line Hep3B.

Protein kinase C suppresses rat organic anion transporting polypeptide 1- and 2-mediated uptake.

Rat oatp1 (Slc21a1) and oatp2 (Slc21a5) transport many structurally unrelated endogenous and exogenous compounds across the sinusoidal membrane of hepatocytes in a sodium-independent manner. There are several potential protein kinase A (PKA) and protein kinase C (PKC) phosphorylation sites in both rat oatp1 and oatp2 proteins, suggesting that PKA and/or PKC may play a role in regulating their function. It is known that the activities of many transporters are subject to the short-term regulation by activation of PKA or PKC, and thus the purpose of the current study was to determine the effect of compounds that activate or inhibit PKA and PKC on the uptake function of rat organic anion transporting protein (oatp)1 and oatp2 when expressed in Xenopus laevis oocytes. In the present investigation, neither the PKA activator N-6-benz-cAMP (0.001-1 mM) nor the PKA inhibitor H7 (0.1-100 microM) affected the uptake mediated by rat oatp1 and oatp2. In contrast, the PKC activator phorbol-12-myristate-13-acetate (PMA) suppressed the uptake mediated by rat oatp1 and oatp2 in a concentration- and time-dependent manner. In addition, pretreatment with bisindolylmaleimide, a specific PKC inhibitor, partially reversed the suppression of PMA on rat oatp1-, and almost completely reversed the suppression of PMA on rat oatp2-mediated uptake. In conclusion, this study indicates that rat oatp1- and oatp2-mediated uptake is subject to the short-term regulation by PKC activation, but not by PKA activation.

Removal of toluene in gas streams by a fibrous-bed trickling filter.

A novel fibrous-bed trickling filter was developed to remove toluene present in contaminated air. Pure culture of Pseudomonas putida F1 was attached on fibrous-bed and utilized toluene as the carbon source. Experimental results indicated the removal efficiency decreased with the increase of inlet concentration. In general, the removal efficiency of toluene was greater than 90% when the inlet loading capacity was below 70 g m-3h-3. The elimination capacity increased with increasing inlet loading capacity, but the increased rate decreased gradually. When the inlet loading capacity increased to 300 g m-3h-1, the elimination capacity could approach to 130 g m-3h-1. The first order kinetics model was useful to describe the removal of toluene in this filter and an excellent linear relationship was found between the apparent first order parameter and inlet concentration (ranging from 1.2 g m-3 to 3.5 g m-3). Also, the performance of fibrous-bed trickling filter was relatively stable during the four-month period of continuous operation. Slight clogging phenomena of filters were observed only under high loading capacity.