Knockdown of ribonuclease inhibitor expression with siRNA in non-invasive bladder cancer cell line BIU-87 promotes growth and metastasis potentials.

Human ribonuclease inhibitor (RI) is a cytoplasmic acidic protein. RI is constructed almost entirely of leucine-rich repeats, which might be involved in some unknown biological functions like other structurally similar proteins besides inhibiting RNase A and angiogenin activities. Our previous experiments demonstrated that up-regulating RI might effectively inhibit some tumor growth and metastasis. However, the down-regulating RI influence on the tumor does not have any report until now, the mechanisms underlying antitumor of RI have not been fully understood. In this study, the efficient RNA interferences of RI were constructed using a plasmid vector and identified with RT-PCR, Western blot and Immunocytochemistry, then were transfected into non-invasive bladder cancer BIU-87 cells. We demonstrated that knockdown RI expression in BIU-87 cells could obviously change the cell morphology, rearrange the microfilaments and extend the lamellipodia, as well as enhance proliferation, increase migration, invasion and matrix metalloprotease level, and also reduce adhesion in vitro. BALB/C nude mice that were injected with the BIU-87 cells transfected RI siRNA showed a significant facilitation of the tumor with heavier tumor weight, higher density of microvessels, lower nm23-H1 and E-Cadherin expressions than those in the control group. Taken together, these experiments suggest that knockdown of RI could promote growth and metastasis potentials of BIU-87 cells. Our present findings reveal the novel mechanism that anti-tumor effect of RI is also involved in suppressing growth and metastasis, besides antiangiogenesis. The results show that RI may be a therapeutic target protein for bladder cancer and may be of biological importance.

Prognostic value of Flotillin-1 expression in patients with solid tumors.

BACKGROUND: In numerous studies, Flotillin-1 was reported to be involved in tumor progression, indicating prognosis in various types of cancer. However, the results were inconsistent. RESULTS: A total of 2473 patients from 13 articles were included. The results indicated that: (1) Patients detected with high expression level of Flotillin-1 protein had a significantly shorter OS (HR =1.64; 95%CI: 1.39-1.88), statistical significance was also observed in subgroup meta-analyses stratified by the cancer type, nationality, detecting method, cutoff value, analysis type, sample size and publication date. (2) Patients with high Flotillin-1 protein expression level had a poorer DFS (HR = 2.49; 95%CI: 1.64-3.35), a worse RFS(HR = 3.26; 95%CI: 1.10-5.43) and a potential shorter PFS(HR = 1.84; 95%CI: 0.81-2.87). (3) The pooled odds ratios (ORs) showed that increased Flotillin-1 level was also related to lymph node metastasis (OR =6.30; 95% CI: 3.15-12.59), distant metastasis (OR =6.02; 95% CI: 1.50-24.06) and more advanced TNM stage (OR =4.69; 95% CI: 2.74-8.03). MATERIALS AND METHODS: A comprehensive retrieval was performed in multiple databases, including PubMed, Embase, Web of Science and CNKI. The relevant articles were screened for investigating the association between increased Flotillin-1 expression level and prognosis. Additionally, clinicopathological features data was also extracted from these studies. CONCLUSIONS: High expression level of Flotillin-1 protein was correlated with poorer clinical outcome. It might serve as a prognostic biomarker and a potential predictive factor of clinicopathology in various tumors. Further well-designed clinical studies should be performed to verify the clinical utility of Flotillin-1 in human solid tumors.

Long non-coding RNA-LET can indicate metastasis and a poor prognosis: a meta-analysis.

INTRODUCTION: Recent studies have reported that long non-coding RNA low expression in tumor (lncRNA-LET) was down-regulated in several cancers. The current meta-analysis aims to determine whether lncRNA-LET can be used as a potential biomarker for metastasis and prognosis. EVIDENCE ACQUISITION: We collected all relevant papers by searching multiple electronic databases (PubMed, EMBASE, Google Scholar, CNKI, Wanfang Database) and explored the association between the expression levels of lncRNA-LET and lymph node metastasis (LNM), distant metastasis (DM) and overall survival (OS). EVIDENCE SYNTHESIS: A total of 383 patients from four studies were finally included. The meta-analysis results showed that LNM occurred more frequently in patients from the lncRNA-LET low expression group than in patients from the lncRNA-LET overexpression group (OR=4.56, 95% CI 2.92-7.12, P<0.00001), and a similar result was observed between lncRNA-LET expression and DM (OR=4.77, 95% CI 2.29-9.94, P<0.0001). Additionally, we found that patients with low expression of lncRNA-LET had a poorer OS than those with lncRNA-LET overexpression (HR=2.39, 95% CI 1.57-3.21, P=0.000). CONCLUSIONS: lncRNA-LET may serve as a common molecular marker for metastasis and prognosis in human cancers.

HOXB7 as a promising molecular marker for metastasis in cancers: a meta-analysis.

Numerous studies on carcinoma have revealed that the expression level of HOXB7 in cancerous tissues was significantly higher than that in noncancerous tissues. Elevated expression of HOXB7 is associated with the susceptibility to lymph node metastasis and distant metastasis in various tumors. In this study, a meta-analysis was performed to involve majority of relevant articles and explore the association of HOXB7 expression level with metastasis in cancer patients. Literature retrieval was conducted by searching in a number of electronic databases (up to December 1, 2015). The meta-analysis was conducted with RevMan 5.3 software and Stata SE12.0. A total of 1,532 patients with carcinoma from 14 studies were included in analysis. The results of meta-analysis demonstrated that lymph node metastasis was observed more frequently in the patients group with high expression level of HOXB7 than in the patients group with low expression level of HOXB7 (odds ratio =2.17, 95% CI: 1.74-2.71, P<0.00001, fixed-effects model). In addition, a similar result was observed in the association between HOXB7 expression and distant metastasis; the odds ratio was 1.77 (95% CI: 1.09-2.88, P=0.02, fixed-effects model). This meta-analysis demonstrated that the overexpression of HOXB7 was significantly associated with metastasis in cancer patients, which may be served as a common molecular marker for indicating cancer metastasis.

Positive association between IL-16 rs1131445 polymorphism and cancer risk: a meta-analysis.

INTRODUCTION: The association between IL-16 rs1131445 polymorphism and cancer risk is not consistent or even contradictory, this meta-analysis aims to investigate the role of IL-16 gene rs1131445 polymorphisms in the risk of cancer. EVIDENCE ACQUISITION: A comprehensive online search was conducted in PubMed, EMBASE and CNKI databases to identify eligible studies. The case-control studies related IL-16 rs1131445 C/T polymorphism with the cancer susceptibility were selected according to the inclusion and exclusion criteria. After extracting the basic data information and quality of literature evaluation, the meta-analysis was performed by using STATA 12.0 software, with calculating odds ratio and 95% confidence interval, and further subgroup analysis, literature publication bias test and sensitivity analysis. EVIDENCE SYNTHESIS: There are totally 1677 cases and 1989 non-tumor controls finally involved. Meta-analysis showed that there are statistical correlations between the IL-16 rs1131445 C/T polymorphism and the cancer risk in Asian populations (TS vs. C, OR=0.80, 95%CI: 0.73-0.88; TT vs. TC, OR=0.75, 95%CI: 0.65-0.87; TT vs. CC, OR=0.69, 95% CI: 0.56-0.84; CC+TC vs. TT, OR=1.36, 95%CI: 1.19-1.55; CC vs. TC+TT, OR=1.27, 95%CI: 1.05-1.53) (all P<0.05). CONCLUSIONS: IL-16 rs1131445 C/T polymorphism is related to the susceptibility to cancer in Asians, suggesting that the C allelic gene of rs1131445 is significantly associated with an increasing cancer risk.

[Comparison of thromboelastography and routine coagulation tests for evaluation of blood coagulation function in patients].

OBJECTIVE: To investigate the correlation and consistency between thromboelastography(TEG) and routine coagulation tests, and to evaluate the value of the two methods in determining the blood coagulation of patients. METHODS: The TEG, routine coagulation tests and platelet counts of 182 patients from the Intensive Care Unit(ICU) and Department of Gastroenterology in our hospital from January to September 2014 were performed and analyzed retrospectively for their correlation, Kappa identity test analysis and chi-square test, and the diagnostic sensitivity and specificity of both methods in the patients with bleeding were evaluated. RESULTS: The TEG R time and PT, R time and APTT showed a linear dependence (P<0.01). The relationship between the TEG K value, α-Angle, MA and Fibrinogen showed a linear dependence (P<0.001). And the relationship between the TEG K value, α-Angle, MA and the platelet count were in a linear dependent way (P<0.001). The Kappa values of the TEG R time with PT and APTT were 0.038 (P>0.05) and 0.061 (P>0.05), respectively. The chi-square test values of the TEG R time with PT and APTT were 35.309 (P<0.001) and 15.848 (P<0.001), respectively. The Fibrinogen and the TEG K value, α-Angle, MA value had statistical significance (P<0.001), with a Kappa value of 0.323, 0.288 and 0.427, respectively. The chi-square test values between Fibrinogen and the TEG K value, α-Angle, MA value were not statistically significant, with X2=1.091 (P=0.296), X2=1.361 (P=0.243), X2=0.108 (P=0.742). The Kappa values of the platelet count and the TEG K value, α-Angle, MA value were 0.379, 0.208 and 0.352, respectively, which were also statistically significant difference (P<0.001). The chi-square test values between the platelet count and the TEG K value, α-Angle, MA value showed a statistically significant difference (P<0.001), with X2=37.5, X2=37.23, X2=26.630. The diagnostic sensitivity of the two methods for the patients with bleeding was less than 50%. CONCLUSION: There was a significant correlation between some TEG parameters and routine coagulation tests, but the consistency is weak. Moreover, the diagnostic sensitivity of two methods in the patients with bleeding is low. It was concluded that the TEG cannot replace the conventional coagulation tests, and the preferable method remains uncertain which could reflect the risk of bleeding.

[Construction of let-7a expression plasmid and its inhibitory effect on k-Ras protein in A549 lung cancer cells].

OBJECTIVE: To elucidate the role of let-7a-mediated gene regulation in the pathogenesis of lung cancer. METHODS: Two template DNA sequences were designed based on hsa-let-7a sequence in miRBase database. The let-7a expression construct and a control plasmid, namely pGenesil-let-7a and pGenesil-control, respectively, were generated by cloning the annealed oligonucleotides into pGenesil-1 and then transfected into A549 cells, which were selected by G418 to establish the lung cancer cell lines stably expressing let-7a-GFP and control-GFP. The living cells were counted by MTT assay and cell growth curves were drawn to analyze the cell proliferation. The k-Ras mRNA level was assessed by semi-quantitative RT-PCR, and the expression of k-Ras protein was determined by Western blotting and immunocytochemistry. RESULTS: The recombinant vectors were verified by sequencing. The cell growth curves indicated that the proliferation of the cells transfected with pGenesil- let-7a were inhibited significantly compared with that of cells transfected with pGenesil-control and A549 cells. Semi- quantitative RT-PCR analysis showed that the levels of k-Ras mRNA almost remained unchanged in cells with or without the treatments. Western blotting and immunocytochemistry demonstrated a significant decrease of k-Ras protein levels in cells transfected with pGenesil-let-7a, but not in cells transfected with pGenesil-control, when compared to A549 cells. CONCLUSION: let-7a over-expression represses the expression of k-Ras protein and significantly inhibits the growth of lung cancer cells.

Let-7a elevates p21(WAF1) levels by targeting of NIRF and suppresses the growth of A549 lung cancer cells.

Down-regulation of let-7 microRNA (miRNA) is a key event in lung cancer. Despite recent advances in survival signaling, the roles of let-7 in the context of lung cancer are not fully clear. In this study, we showed that let-7a, a member of let-7 family, negatively regulated the expression of NIRF through NIRF 3' UTR. We also showed that NIRF was required for the let-7a-mediated elevation of p21(WAF1). These findings suggest that growth-inhibitory effect of let-7a on the A549 cells in vitro and in vivo may be explained in part by le-7a-induced suppression of NIRF and elevation of p21(WAF1). This work reveals a novel regulatory mechanism for let-7a in the control of cellular proliferation and lung carcinogenesis.

Research on the antitumor effect of ginsenoside Rg3 in B16 melanoma cells.

Ginsenoside Rg3 is an effective chemical component extracted from the red Panix. The experiment demonstrated that it might effectively inhibit proliferation and metastasis of tumor cells. The exact molecular mechanism of Rg3 remains unclear so far. To further explore the antitumor function of Rg3, we investigated the in-vitro and in-vivo activity of Rg3 in the treatment of B16 melanoma cells, derived from C57BL/6 mouse, capable of forming tumor colonies in the lungs following intravenous injection. Cell proliferation was measured by 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide assay. Morphological changes of cells were observed by staining with Giesma and Hoechst 33258. Cell cycle and apoptosis rate were analyzed by flow cytometry. The expression of caspase-3 and bcl-2 in cells was detected by immunocytochemistry and western blot analysis. We found that Rg3 could inhibit cell proliferation, regulate cell cycle, and induce cell apoptosis in vitro. B16 melanoma-bearing mice were used to evaluate in vivo the antitumor activity of Rg3. Mice that were injected with Rg3 showed significant inhibition of the tumor metastasis with lighter lung weight, lower density of microvessels, fewer metastasis nodules, and longer survival time than those in the control group (P<0.001). In conclusion, the results reveal that antitumor metastasis of Rg3 is also associated with inducing apoptosis, regulating cell cycle, and blocking angiogenesis in addition to inhibiting proliferation. This research might supply valuable data for chemotherapy with Rg3 in melanoma. Rg3 would turn out to be an anticancer drug with promising prospects.

[PET/CT-based classification of delayed radiation encephalopathy following radiotherapy for nasopharyngeal carcinoma].

OBJECTIVE: To investigate positron-emission tomography-computed tomography (PET/CT) findings of radiation encephalopathy (RE) following radiotherapy for nasopharyngeal carcinoma (NPC), observe the metabolic changes of the compromised brain tissues, and postulate the clinical classification of RE to provide reference for its diagnosis. METHODS: This study included 53 pathologically confirmed NPC patients who received previous radical radiotherapy, and the diagnosis of RE was established according to the clinical manifestations and CT/PET findings. All the patients underwent PET/CT whole-body and head scans, and the image data were evaluated along with the clinical data of the patients. RESULTS: RE most frequently involved the lateral or bilateral inferior temporal lobes. PET identified hypometabolic changes in the bilateral temporal lobes of 35 patients (70 lobes) and in the lateral temporal lobe of 18 patients (18 lobes). According to the PET/CT findings, the lesions were classified into 3 types, namely the oedema type (56 temporal lobes), liquefactive necrosis type (10 temporal lobes), and atrophic calcification type (22 temporal lobes). One patient with oedema type lesion received neurotrophic treatment and recovered completely with normal brain tissue density and metabolism, but the oedema type lesions in 2 patients progressed into to atrophic calcification type; the liquefactive necrotic lesions in another 2 patients also progressed into atrophic calcification type. CONCLUSION: RE patients exhibit significant hypometabolic changes in the inferior temporal lobe on PET. According to the findings by PET/CT, RE can be classified into the oedema type, liquefactive necrosis type, and atrophic calcification type, and lesions of the former two types may progress into the third type.