非编码RNA在喉癌发生中的作用及其临床意义

喉癌是头颈部最常见的恶性肿瘤之一,喉癌患者吞咽、呼吸及发音功能均可受到严重影响,患者经受“有苦难言”的折磨,生活质量极差。因此,揭示喉癌发病的分子机制,以期提高喉癌早期诊断水平和防治效果一直是该研究领域的热点之一。近年来,随着基因测序技术、转录组学技术和生物信息学技术等分子生物学技术的应用,越来越多的与喉癌发生相关的非编码RNA先后被发现。实验证明,一些短链非编码RNA（如：上调的miR-16、miR-21、miR．106b和miR-1297,下调的let．7a、miR．1、miR．24、miR．34a／c、miR-137、miR-203和miR-206）以及长链非编码RNAf如：H19、HOTAIR和MALAT-1）均被发现参与了喉癌的发生、发展过程,它们发挥着促癌或者抑癌作用,影响细胞的增殖、浸润、转移和凋亡。这些非编码RNA将有可能为喉癌诊断和治疗分别提供新的标志物和治疗靶点。     

miR‐203 inhibits proliferation of HCC cells by targeting survivin

To validate whether down‐regulation of microRNA‐203 (miR‐203) in hepatocellular carcinoma (HCC) is involved in HCC progression by targeting survivin. MiR‐203 mimics was transfected into HepG2 cells to enhance miR‐203 expression, and miR‐203 inhibitor was transfected into HepG2 cells to inhibit miR‐203 expression. The effect of up‐regulation and down‐regulation of miR‐203 on survivin expression of HepG2 cells was evaluated using Western blot assay. The effect of miR‐203 or survivin expression on the proliferation of HepG2 cells was detected using the CKK‐8 assay. Over‐expression of miR‐203 significantly inhibited the expression of survivin in HepG2 cells (p < 0·05), and down‐expression of miR‐203 significantly promoted the expression of survivin in HepG2 cells (p < 0·05). Both over‐expression of miR‐203 and down‐regulation of survivin suppressed proliferation of HepG2 cells significantly compared with negative control. Low expression of miR‐203 contributes to the progression of HCC via targeting survivin. Copyright © 2012 John Wiley & Sons, Ltd.     

DGCR5 promotes cancer stem cell-like properties of radioresistant laryngeal carcinoma cells by sponging miR-506 via Wnt pathway

Cancer stem cells (CSCs) have been recognized as the significant cause of tumor recurrence. Long noncoding RNAs (lncRNAs) are involved in various cancers, including human laryngeal cancer. So far the correlation between lncRNA DiGeorge syndrome critical region gene 5 (DGCR5) and CSC-like properties in human laryngeal cancer remains barely known. In our current study, two human larynx squamous carcinoma cell lines (Hep-2 and Hep-2R) with different radio sensitivities were cultured. Interestingly, CSC-like phenotypes were much more enriched in Hep-2R cells. We found that DGCR5 was upregulated and microRNA-506 (miR-506) was downregulated in Hep-2R cells. In addition, silence of DGCR5 could inhibit the stemness and enhance the radiosensitivity of Hep-2R cells. Meanwhile, overexpression of miR-506 also suppressed the CSC-like traits and the radiosensitivity was increased significantly. In addition, miR-506 was predicted as target of DGCR5 and the correlation between them was validated in our study. Finally, we observed that Wnt pathway exerted a significant role in human laryngeal CSCs and DGCR5 inhibition could repress Wnt signaling activity by sponging miR-506. In vivo assays were performed and we found that DCGR5 depressed stemness of human laryngeal cancer cells through modulating miR-506 and Wnt signaling pathway. Taken these together, we reported that DGCR5 induced CSC-like properties by sponging miR-506 through activating Wnt in human laryngeal carcinoma cells.     

Knockdown of DGCR5 enhances the radiosensitivity of human laryngeal carcinoma cells via inducing miR-195

Long noncoding RNAs (lncRNAs) exert critical roles in the development of various cancers, including human laryngeal cancer. Radioresistance contributes to the predominant causes of laryngeal cancer recurrence after radiotherapy. The aim of our study was to investigate the association of dysregulated lncRNA and radiation resistance in human larynx squamous carcinoma. Here, we investigated the biological roles of lncRNA DiGeorge syndrome critical region gene 5 (DGCR5) in radioresistance of human laryngeal cancer. Two human larynx squamous carcinoma cell lines (Hep-2 and Hep-2R), with different radiosensitivities in vitro were used in the present study. We observed that DGCR5 was significantly upregulated in Hep-2R cells. Inhibition of DGCR5 by LV-shDGCR5 transfection restrained Hep-2R cell proliferation and sensitized cells to radiation. Reversely, overexpression of DGCR5 exhibited an opposite phenomenon in vitro. In addition, microRNA (miR)-195 was predicted as a direct downstream target of DGCR5. Dual-luciferase reporter and RNA immunoprecipitation assays verified the direct interaction between them. Meanwhile, miR-195 was observed to be reduced in Hep-2R cells and miR-195 mimics repressed Hep-2 cell growth. Moreover, radiosensitivity of Hep-2R cells was greatly enhanced by overexpression of miR-195, which could be reversed by upregulation of DGCR5. Finally, in vivo experiments were used to validate that knockdown of DGCR5 suppressed laryngeal carcinoma via targeting miR-195. In conclusion, we indicated that DGCR5 could contribute to the radioresistance of human laryngeal carcinoma cells via sponging miR-195.     

microRNA-203 suppresses bladder cancer development by repressing bcl-w expression

It is increasingly clear that microRNAs (miRNAs) play an important role in many diseases, including tumorigenesis. However, the mechanisms by which miRNAs regulate bladder cancer development remain poorly understood. Here, we evaluated the expression of microRNA-203 (miR-203) in bladder cancer tissues using real-time PCR, and defined the target genes and biologically functional effect using luciferase reporter assay, flow cytometry and western blot analysis. We first verified that the expression of miR-203 was decreased in bladder cancer tissues. Moreover, ectopic expression of miR-203 promoted the apoptosis of human bladder cancer cell lines and inhibited cell proliferation, whereas its depletion increased cell growth. We further verified that miR-203 directly targeted 3'-untranslated region of the bcl-w gene, and decreased its expression in vitro and in vivo. Western blot analysis also showed that the expression level of miR-203 was negatively correlated with bcl-w level in tumor tissues. These data suggest an important role for miR-203 in the molecular etiology of bladder cancer and implicate the potential application of miR-203 in bladder cancer therapy.     

miR-203在肝细胞肝癌中的表达及其对肿瘤细胞增殖及侵袭能力的影响

目的:探讨miR-203在肝细胞肝癌(Hepatocellular carcinoma,HCC)患者血浆中的表达及其对肝癌细胞增殖和侵袭能力的影响。方法:选择2013年5月-2015年7月在我院确诊为肝细胞肝癌的患者57例作为研究对象,另选取同期在我院接受健康体检的志愿者49例作为对照组。采用实时荧光定量RT-PCR法检测两组血浆中miR-203的表达水平;将过表达的miR-203转染至人肝癌细胞系Hep 3B和JHH-1;采用CCK-8法检测miR-203对肝癌细胞增殖能力的影响;采用transwell法检测miR-203对肝癌细胞迁移及侵袭能力的影响。结果:miR-203在肝癌患者血浆中的表达水平显著低于对照组,差异具有统计学意义(P0.01);与未转染的肝癌细胞比较,Hep 3B和JHH-1细胞转染miR-203mimic后,肿瘤细胞的增殖、迁移及侵袭能力受到明显抑制,差异具有统计学意义(P0.05)。结论:miR-203在肝细胞肝癌中呈低表达,而过表达miR-203能够抑制肿瘤细胞的增殖及侵袭能力,提示miR-203可以作为一种新的肿瘤抑制性micro RNA。     

MicroRNA-203 reinforces stemness properties in melanoma and augments tumorigenesis in vivo

One of the challenges encountered in microRNA (miRNA) studies is to observe their dual role in different conditions and cells. This leads to a tougher prediction of their behavior as gene expression regulators. miR-203 has been identified to play a negative role in the progression of malignant melanoma; however, it has been reported, with dual effect, as both an oncomiR and tumor suppressor miRNA in some malignancies, such as breast cancer, meanwhile, the role of miR-203 in melanoma stem cells or even metastatic cells is unclear. In the present study, after observation of upregulation of miR-203 in melanoma patient's serum and also melanospheres as cancer stem cells model, we examined its overexpression on the stemness potential and migration ability of melanoma cells. Our data demonstrated that the increased miR-203 level was significantly associated with significant increase in the ability of proliferation, colony and spheres formation, migration, and tumorigenesis in A375 and NA8 cells. All of these changes were associated with enhancement of BRAF, several epithelial to mesenchymal transition factors, and stemness genes. In conclusion, our results clearly determined that miR-203 could be down-regulateddownregulated in melanoma tissues but be overexpressed in melanoma stem cells. It has an important role as oncomiR and promote repopulation, tumorigenicity, self-renewal, and migration. Therefore, we suggested overexpression of miR-203 as biomarker for early detection of metastasis. However, more studies are needed to validate our data.     

miR-126增加非小细胞肺癌细胞A549对顺铂的敏感性

miR-126在多种恶性肿瘤中存在表达下调并显示抑癌基因的功能,然而其在肿瘤敏感性中的作用仍不明确.为了探讨miR-126在非小细胞肺癌细胞A549对顺式铂氨(cis-diammine dichloroplatoum, cisplatin, CDDP)敏感性中的作用及可能机制,本研究用MTS法检测非小细胞肺癌细胞A549及其衍生的CDDP耐受细胞A549/DDP对CDDP的敏感性.结果表明,A549/DDP细胞对CDDP的耐受性是A549细胞的4.05倍(P=0.0078)|用qRT-PCR检测发现,相比于A549细胞,A549/DDP细胞中miR-126的表达下调了8.45倍(P=0.0063),而survivin和Bcl-2的表达明显上调|通过MTS、qRT-PCR及Western印迹实验发现,miR-126 mimics使A549/DDP细胞中miR-126的表达上调了12.63倍(P=0.0013),并明显增加A549/DDP细胞对CDDP的敏感性及下调survivin和Bcl-2的表达;相反,miR-126 inhibitor能明显增加A549细胞对CDDP的耐受性及增加survivin和Bcl-2的表达.本研究结果提示,miR-126在非小细胞肺癌CDDP耐受细胞中的表达下调,上调miR-126的表达能增加耐药细胞对CDDP的敏感性. miR-126是逆转肺癌CDDP耐受的可能潜在靶标.     

Significance and relationship between DJ-1 gene and survivin gene expression in laryngeal squamous cell carcinoma

This study aimed at exploring the correlation between DJ-1 gene and survivin gene in laryngeal squamous cell carcinoma by analyzing their gene expression levels and their relationship with clinicopathologic parameters. The expression of DJ-1 gene and survivin gene in 82 laryngeal carcinoma tissues from patients and 82 negative surgical margin tissue samples were detected by immunohistochemistry, respectively. The correlation of their expression levels and patients'' clinical parameters were then analyzed by Pearson correlation analysis. The positive detection rates of DJ-1 and survivin in laryngeal carcinoma tissues were 71.95% and 60.98%, which were higher than those of the normal control that were 29.27% and 0.00%, respectively (P<0.01). The positive detection rates of DJ-1 and survivin were found associated with tumor stages (P<0.05), but not with lymph node metastasis. The DJ-1 gene expression level was related to cell differentiation (P<0.05). Finally, a positive correlation between DJ-1 and survivin gene expression in laryngeal carcinoma was found. The overall survival rate of patients was 51.2%, and disease-free survival (DFS) was 39.0%. DFS in DJ-1 negative-expression group was 87.0%, and 20.3% in DJ-1 positive-expression group. The negative expression of DJ-1 was associated with a shorter mean patient DFS time (44.643±1.417 months), whereas positive expression of DJ-1 was associated with a longer mean DSF time (25.943±;1.297 months). DJ-1 and survivin play a vital role in the occurrence and development of laryngeal carcinoma. DJ-1 may promote the carcinogenesis of laryngeal cells by up-regulating the survivin gene expression.Key words: laryngeal carcinoma, DJ-1 gene, survivin gene, diagnosis.     

Significance and relationship between DJ-1 gene and surviving gene expression in laryngeal carcinoma

This study aimed at exploring the correlation between DJ-1 gene and survivin gene in laryngeal squamous cell carcinoma by analyzing their gene expression levels and their relationship with clinicopathologic parameters. The expression of DJ-1 gene and survivin gene in 82 laryngeal carcinoma tissues from patients and 82 negative surgical margin tissue samples were detected by immunohistochemistry, respectively. The correlation of their expression levels and patients’ clinical parameters were then analyzed by Pearson correlation analysis. The positive detection rates of DJ-1 and survivin in laryngeal carcinoma tissues were 71.95% and 60.98%, which were higher than those of the normal control that were 29.27% and 0.00%, respectively (P<0.01). The positive detection rates of DJ-1 and survivin were found associated with tumor stages (P<0.05), but not with lymph node metastasis. The DJ-1 gene expression level was related to cell differentiation (P<0.05). Finally, a positive correlation between DJ-1 and survivin gene expression in laryngeal carcinoma was found. The overall survival rate of patients was 51.2%, and disease-free survival (DFS) was 39.0%. DFS in DJ-1 negative-expression group was 87.0%, and 20.3% in DJ-1 positive-expression group. The negative expression of DJ-1 was associated with a shorter mean patient DFS time (44.643±1.417 months), whereas positive expression of DJ-1 was associated with a longer mean DSF time (25.943±1.297 months). DJ-1 and survivin play a vital role in the occurrence and development of laryngeal carcinoma. DJ-1 may promote the carcinogenesis of laryngeal cells by up-regulating the survivin gene expression.     

Diagnostic and mechanistic values of microRNA-130a and microRNA-203 in patients with papillary thyroid carcinoma

This research was determined to unearth the diagnostic values and the effects of microRNA (miR)-130a and miR-203 on cell proliferation and apoptosis of papillary thyroid carcinoma (PTC). Expression of miR-130a and miR-203 were evaluated and were subjected to correlation analysis. The diagnostic values of miR-130a and miR-203 and their associations with clinicopathological characteristics of patients with PTC were measured. The expression levels of miR-130a and miR-203 in K1, IHH4, TPC-1, and BCPAP cells together with Nthy-ori 3-1 cells were measured. Cells were transfected with miR-130a mimics, miR-203 mimics, and coordinate of miR-130a mimics and miR-203 mimics. Cell growth, colony formation, and apoptosis were detected by cell counting kit-8 (CCK-8) assay, colony formation assay, and flow cytometry. PTC tissues had decreased miR-130a and miR-203 relative to adjacent normal tissues and normal thyroid tissue (both P < .05). miR-130a was in positive correlation with miR-203 (r = 0.754, P < .01). miR-130a was related with tumor infiltration and tumor stage while miR-203 was implicated in tumor stage and lymph-node metastasis. The area under the curve (AUC), sensitivity, as well as specificity for miR-130 in predicting PTC was 0.839, 74.5%, and 85.0% and those for miR-203 were 0.818, 73.7%, and 84.0%, respectively. PTC cells had lower expression of miR-130a and miR-203 than that in Nthy-ori 3-1 cells. After transfected miR-130a and miR-203 mimics in BCPAP and TPC-1 cells, both cells had increased miR-130a and miR-203, promoted cell apoptosis rate and decreased cell growth rate, and colony formation ability. After coordinately transfected with miR-130a mimics and miR-203 mimics, the cell growth and colony formation ability of PTC cells were restrained, and apoptosis of PTC cells was elevated (all P < .05). This study highlights that miR-130a and miR-203 have satisfactory diagnostic value in PTC and upregulated miR-130a and miR-203 can inhibit PTC cell growth and promote cell apoptosis.     

Upregulation of microRNA-141 suppresses epithelial-mesenchymal transition and lymph node metastasis in laryngeal cancer through HOXC6-dependent TGF-β signaling pathway

Laryngeal cancer is one of the most malignant cancers among the head and neck malignant tumors. Abnormal expression of microRNAs (miRNAs) contributes to cancer development through regulating proliferation and apoptosis of cancer cells. In this study, we aim to explore the roles of microRNA-141 (miR-141), Homeobox C6 (HOXC6) and TGF-β signaling pathway in epithelial-mesenchymal transition (EMT) and lymph node metastasis in laryngeal cancer. Initially, we identified differentially expressed genes in laryngeal cancer, among which HOXC6 was identified. Then the target miRNA of HOXC6 was predicted and verified. Next, expression of miR-141, HOXC6, TGF-β1, Smad3, Vimentin and Snail in cancer tissues was detected. Then, AMC-HN-8 cells were transfected with miR-141 mimic, miR-141 inhibitor and HOXC6-siRNA to investigate specific role of miR-141, HOXC6 and TGF-β signaling pathway in laryngeal cancer in vivo and in vitro. Our results showed that HOXC6 was a target gene of miR-141, which was downregulated in laryngeal cancer. Besides, overexpression of miR-141 could downregulate HOXC6 and inhibit the TGF-β signaling pathway. Upregulation of miR-141 or silencing of HOXC6 can repress EMT, viability, migration and invasion abilities of laryngeal cancer cells. In addition, upregulation of miR-141 inhibited the tumor growth and lymph node metastasis in vivo. In summary, our findings demonstrated that upregulated miR-141 decreased HOXC6 expression, and inhibited the TGF-β signaling pathway, EMT and lymph node metastasis in laryngeal cancer, which is of clinical significance in the treatment of laryngeal cancer.