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ABSTRACT: MiR-214 has been shown to inhibit cell growth, migration and invasion. Here we demonstrate that ectopic expression of miR-214 reduces cell survival, induces apoptosis and enhances sensitivity to cisplatin through directly inhibiting Bcl2l2 expression in cervical cancer HeLa and C-33A cells. Further analysis reveals that apoptosis induced by miR-214 is correlated with increased expression of Bax, caspase-9, caspase-8 and caspase-3. Moreover, we show that miR-214 is regulated by DNA methylation and histone deacetylation. Taken together, these data suggest that miR-214 might be a candidate target for the development of novel therapeutic strategies to treat cervical cancer.
FEBS letters 01/2013; · 3.54 Impact Factor
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ABSTRACT: MicroRNAs are a class of small endogenous noncoding RNAs that function as post-transcriptional regulators. Tumor protein p53-induced nuclear protein 1 (TP53INP1) is a p53 target gene and is a major player in the stress response. Here, we identified TP53INP1 as a target of miR-17-5p. miR-17-5p suppressed cell growth and promoted apoptosis of cervical cancer cells, whereas the effects of TP53INP1 were opposite, and ectopic expression of TP53INP1 counteracted the suppression of cell growth caused by miR-17-5p. The same correlations between miR-17-5p and TP53INP1 were observed in cervical cancer tissues. Together, these results indicated that miR-17-5p functions as a tumor suppressor in cervical cancer cells by targeting TP53INP1.
International Union of Biochemistry and Molecular Biology Life 06/2012; 64(8):697-704. · 3.51 Impact Factor
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ABSTRACT: MicroRNAs (miRNAs) play a key role in the regulation of gene expression. In this study, we demonstrate that microRNA-19a and -19b (miR-19a/b) are highly expressed in human cervical cancer cells and are involved in maintaining the malignant phenotypes of HeLa and C33A cells. Up-regulation of miR-19a and miR-19b promoted cell growth and invasion, whereas knockdown of miR-19a and miR-19b yielded the reverse phenotype. CUL5 was identified as a novel target gene of both miR-19a and miR-19b. CUL5 ectopic over-expression without its 3' untranslated region (UTR) abolished the effect of miR-19a/b on HeLa and C33A cell proliferation and invasion. These results indicated that miR-19a/b directly and negatively regulate CUL5 expression in cervical cancer cells, highlighting the importance of miR-19a and miR-19b and their target genes in tumorigenesis.
Cancer letters 05/2012; 322(2):148-58. · 4.86 Impact Factor
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ABSTRACT: Recently, PIWIL4 has been identified as a functional protein involved in tumorigenesis. Cervical cancer is the second most prevalent form of cancer worldwide. The relationship between PIWIL4 and cervical cancer is still unknown. Here, we found that PIWIL4 is up-regulated in human cervical cancer tissues in comparison to adjacent normal tissues, and it promotes cell growth and proliferation by inhibiting apoptosis through the p14ARF/p53 pathway. PIWIL4 can also promote the invasion of cervical cancer cells. These results suggest that PIWIL4 might play an oncogenic role in cervical cancer and be useful as a new therapeutic target in the future.
FEBS letters 03/2012; 586(9):1356-62. · 3.54 Impact Factor
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ABSTRACT: miR-20a is an important member of the miR-17-92 cluster, and its real function in cervical cancer cells is unknown. Our study demonstrated that miR-20a was upregulated in cervical cancer tissues. Overexpression of miR-20a in cervical cancer-derived cell lines, HeLa and C-33A, enhanced long-term cellular proliferation, migration and invasion, whereas inhibition of miR-20a suppressed those functions. We also confirmed that oncogenic TNKS2 is directly upregulated by miR-20a. Furthermore, suppression of TNKS2 expression could inhibit colony formation, migration and invasion of cervical cancer cells. Therefore, we concluded that miR-20a can promote migration and invasion of cervical cancer cells through the upregulation of TNKS2.
FEBS letters 03/2012; 586(6):897-904. · 3.54 Impact Factor
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ABSTRACT: MicroRNAs (miRNAs) are an evolutionarily conserved class of endogenous, non-coding RNAs that modulate gene expression at the post-transcriptional level and are involved in tumorigenesis. In this study, we demonstrate that miR-1 suppresses the potential for growth, migration and invasion in the HEp2 cell line. Furthermore, we validate that FN1 is a direct target gene for miR-1 via fluorescent reporter assay and is negatively regulated by miR-1. Moreover, the knockdown of FN1 has the same phenotypic effects as the overexpression of miR-1. Taken together, our results provide evidence that miR-1 may play a role as a tumor suppressor gene in laryngeal carcinoma.
FEBS letters 09/2011; 585(20):3263-9. · 3.54 Impact Factor
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ABSTRACT: MicroRNAs (miRNAs) are small non-coding RNAs that regulate the translation of target mRNA transcripts. In this study, we demonstrated that miR-519d was downregulated in human HCC and could suppress growth of the human HCC cell line QGY-7703. Bioinformatics analysis indicated that MKi67 was a putative target of miR-519d. In an EGFP reporter system, we confirmed that MKi67 was a direct target gene of miR-519d. Furthermore, knockdown of MKi67 inhibited QGY-7703 cell growth. These findings indicate that miR-519d targets the MKi67 transcript and suppresses HCC cell growth, suggesting that miR-519d has a tumor suppressive role in human HCC pathogenesis.
Cancer letters 08/2011; 307(2):182-90. · 4.86 Impact Factor
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ABSTRACT: The posttranscriptional regulation of miRNAs is important for organism development. To investigate the role of miRNAs in angiogenesis, we performed a loss-of-function screening assay in human umbilical vein endothelial cells (HUVECs) and found that knockdown of 7 miRNAs (miR-95a, miR-126, miR-129, miR-137, miR-139, miR-200a, and miR-335) significantly suppressed cell viability. As miR-200a was highly expressed in HUVECs, blocking endogenous miR-200a using 2'-OMe antisense oligonucleotide (ASOs) resulted in a decrease of cell viability and migration. Bioinformatics analysis indicates the 3' untranslated region (UTR) of thrombospondin-1 (THBS1) has a putative binding site for miR-200a. MiR-200a can directly bind to THBS1 3'UTR and negatively regulate THBS1 expression. The identification of endothelial cells (ECs) related miRNA and its target gene may gain new insight into the mechanism of angiogenesis.
International Union of Biochemistry and Molecular Biology Life 07/2011; 63(7):553-9. · 3.51 Impact Factor
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ABSTRACT: MicroRNAs are a class of noncoding RNAs that are ~22 nucleotides in length. MicroRNAs have been shown to play important roles in cell differentiation and in cancer. Recently, studies have shown that miR-372 is tumorigenic in human reproductive system cancers. However, we provide evidence that miR-372 acts as a tumor suppressor gene in cervical carcinoma. miR-372 was found down-regulated in cervical carcinoma tissues as compared with adjacent normal cervical tissues. Growth curve and FACS assays indicated that ectopic expression of miR-372 suppressed cell growth and induced arrest in the S/G₂ phases of cell cycle in HeLa cells. We used bioinformatic predictions to determine that CDK2 and cyclin A1 were possible targets of miR-372 and confirmed this prediction using a fluorescent reporter assay. Taken together, these findings indicate that an anti-oncogenic role of miR-372 may be through control of cell growth and cell cycle progression by down-regulating the cell cycle genes CDK2 and cyclin A1.
Journal of Biological Chemistry 06/2011; 286(29):25556-63. · 4.77 Impact Factor
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ABSTRACT: RAC1 regulates a diverse array of cellular events including migration and invasion. MicroRNAs (miRNAs) have a key role in the regulation of gene expression. In this study, we demonstrated that microRNA-142-3p (miR-142-3p) acted as a negative regulator of human RAC1. Overexpression of miR-142-3p decreased RAC1 mRNA and protein levels. Moreover, the overexpression of miR-142-3p suppressed, while blocking of miR-142-3p increased colony formation, migration and invasion in hepatocellular carcinoma (HCC) cell lines (QGY-7703 and SMMC-7721). RAC1 overexpression without the 3'untranslated region abolished the effect of miR-142-3p in the QGY-7703 and SMMC-7721 cells. These results demonstrated that miR-142-3p directly and negatively regulates RAC1 in HCC cells, which highlights the importance of miRNAs in tumorigenesis.
FEBS letters 05/2011; 585(9):1322-30. · 3.54 Impact Factor
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ABSTRACT: To investigate the role of the peptide Pro-His-Ser-Arg-Asn (PHSRN) in cell adhesion and growth, PHSRN- and Gly-Arg-Gly-Asp-Ser (GRGDS)-containing polymers (P-PN5 and P-GS5, respectively) were synthesized by modification of poly(D,L-lactide-co-beta-malic acid) (PLMA) with the corresponding peptides. The cell affinities of the modified polymers were evaluated by adhesion and proliferation experiments with human umbilical vein endothelial cells (HUVECs). The results showed that P-PN5 had comparable ability to that of P-GS5 in supporting HUVEC adhesion and growth. Furthermore, the integrin-mediated mechanism of cell-substrate interaction was investigated. The results showed that P-PN5 had similar binding affinity and binding strength towards α(5)β(1) compared to those of P-GS5. The findings suggest that PHSRN is capable of mediating the adhesion and growth of HUVECs independently and that PHSRN-modified polymers might be used as biologically compatible materials.
Colloids and surfaces. B, Biointerfaces 05/2011; 84(1):6-12. · 2.60 Impact Factor
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ABSTRACT: MicroRNAs are a class of small noncoding RNAs that function as key regulators of gene expression at the post-transcriptional level. Recently, microRNA-373 (miR-373) has been found to function as an oncogene in testicular germ cell tumors. In our study, we found that miR-373 is upregulated in human hepatocellular carcinoma (HCC) tissues as compared with adjacent normal tissues, and promotes the proliferation of the HCC cell lines HepG2 and QGY-7703 by regulating the transition between G(1)-phase and S-phase. The gene encoding the protein phosphatase 6 catalytic subunit (PPP6C ), a negative cell cycle regulator, was identified as a direct target gene of miR-373 by use of a fluorescent reporter assay. The mRNA and protein levels of PPP6C were both inversely correlated with the miR-373 expression level. Overexpression of PPP6C abolished the regulation of cell cycle and cell growth exercised by miR-373 in HepG2 cells. These results indicate that miR-373 plays an important role in the pathogenesis of HCC, and may be a new biomarker in HCC. Our results demonstrate that miR-373 can regulate cell cycle progression by targeting PPP6C transcripts and promotes the growth activity of HCC cells in vitro. The downregulation of PPP6C by miR-373 may explain why the expression of miR-373 can promote HCC cell proliferation.
FEBS Journal 04/2011; 278(12):2044-54. · 3.79 Impact Factor
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ABSTRACT: Chemotherapy is an important treatment for colorectal adenocarcinoma cancer; however, colorectal adenocarcinoma cells often develop resistance to chemotherapeutic drugs, leading to relapse and poor patient prognosis. The development of drug resistance is often a multifactor process, which involved several genes and cellular mechanisms. microRNAs are endogenous small non-coding RNAs that negatively regulate gene expression at the post-transcriptional level. In the present study, we investigated the possible role of microRNAs in regulating drug sensitivity of colorectal adenocarcinoma cells SW620 and SW480. Using microRNA expression arrays and quantitative reverse transcriptase (RT)-PCR, we found that SW620 cells exhibited elevated miR-20a expression compared with SW480 cells. In addition, these two cell lines displayed different sensitivities to the chemotherapeutic drugs fluorouracil, oxaliplatin, and teniposide. Modulation of miR-20a altered the sensitivity of SW620 and SW480 cells to these drugs; knockdown of miR-20a sensitized SW620 cells to chemotherapeutic agents, whereas overexpression of miR-20a in SW480 cells resulted in chemoresistance. Endogenous BNIP2 mRNA and BNIP2 protein levels were inversely related to miR-20a levels as detected by quantitative RT-PCR and western blot analysis. Fluorescence reporter assays showed a direct interaction between miR-20a and the BNIP2 3'UTR. Taken together, our findings suggested that miR-20a may play a role in colorectal adenocarcinoma cancer cell drug resistance and may be a therapeutic target against chemotherapy drug resistance in colorectal adenocarcinoma.
Acta Biochimica et Biophysica Sinica 03/2011; 43(3):217-25. · 1.38 Impact Factor
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ABSTRACT: MicroRNAs are small noncoding RNAs that negatively regulate target mRNAs at the posttranscription level. Here, we show that miR-16 is upregulated in human laryngeal carcinoma tissues. Inhibition of miR-16 in HEp-2 laryngeal cancer cell line could suppress cell migration and enhance cell-cell adhesion. Subsequently, zyxin, whose expression is negatively regulated by miR-16, is confirmed to be a direct target gene of miR-16. Furthermore, overexpression of zyxin could also restrain cell movement and enhance cell-cell adhesion. The study of miR-16 and its target zyxin will shed light on diagnosis and therapy of laryngeal cancer.
International Union of Biochemistry and Molecular Biology Life 02/2011; 63(2):101-8. · 3.51 Impact Factor
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Ran Qiang,
Fang Wang,
Li-Ying Shi, Min Liu,
Shuang Chen,
Hai-Ying Wan,
Yi-Xuan Li,
Xin Li,
Song-Yuan Gao,
Bao-Cun Sun,
Hua Tang
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ABSTRACT: Plexin-B1, the receptor for Sema4D, has been reported to trigger multiple and sometimes opposing cellular responses in various types of tumor cells. It has been implicated in the regulation of tumor-cell survival, proliferation, angiogenesis, invasion and metastasis. However, the plexin-B1 gene expression and its regulatory mechanism in cervical cancer remain unclear. The present study shows that plexin-B1 is over-expressed in cervical tumor tissues compared to normal cervical tissues by immunohistochemistry, Western blotting and quantitative RT-PCR. The expression of plexin-B1 is significantly associated with cervical tumor metastasis and invasion according to the analysis of the clinicopathologic data. Plexin-B1 also promotes proliferation, migration and invasion in human cervical cancer HeLa cells. We also found that the plexin-B1 levels are inversely correlated with miR-214 amounts in both cervical cancer tissues and HeLa cells. And miR-214 expression level is also associated with metastasis and invasion of cervical tumor. Furthermore, we demonstrate that plexin-B1 is inhibited by miR-214 through a miR-214 binding site within the 3'UTR of plexin-B1 in HeLa cells. Ectopic expression of miR-214 could inhibit the proliferation capacity, migration and invasion ability of HeLa cells. Our findings suggest that plexin-B1, a target of miR-214, may function as an oncogene in human cervical cancer HeLa cells.
The international journal of biochemistry & cell biology 01/2011; 43(4):632-41. · 4.89 Impact Factor
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ABSTRACT: Accumulating evidence suggests that microRNAs (miRNAs) control the replication of both RNA and DNA viruses. In order to determine whether host-encoded miRNAs affect hepatitis B virus (HBV) replication, antisense oligonucleotides (ASOs) of 328 identified human miRNAs were transfected into HepG2 2.2.15 cells, respectively. ELISA and MTS assay were used to measure the expression level of HBV S protein (HBsAg), HBV e antigen (HBeAg) and cell proliferation. Compared to experimental controls, miR-199a-3p and miR-210 efficiently reduced HBsAg expression without affecting HepG2 2.2.15 cell proliferation. Quantification of HBV DNA by real-time PCR showed that both miRNAs suppressed viral replication. Bioinformatics analysis indicated a putative binding site for miR-199a-3p in the HBsAg coding region and a putative binding site for miR-210 in the HBV pre-S1 region. The direct effect of miRNAs on the target region in HBV transcripts was validated by a fluorescent reporter assay, and the suppression of HBs gene expression by both miRNAs was measured by real-time PCR and Western blot. These results suggest that up-regulation of miR-199a-3p and miR-210 in HepG2 2.2.15 cells compared to HepG2 cells may play a role in regulating HBV replication and maintenance of a suitable level of virion production in persistent infection by targeting crucial HBV genes.
Antiviral research 11/2010; 88(2):169-75. · 3.61 Impact Factor
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ABSTRACT: MicroRNAs are an evolutionarily conserved class of endogenous noncoding RNAs that modulate gene expression at the post-transcriptional level. Recently, microRNA-23a (miR-23a) has been found to function as a growth-promoting and antiapoptotic factor in hepatocellular carcinoma cells. Our previous study showed that miR-23a was significantly upregulated in gastric adenocarcinoma tissues. In this study, we found that miR-23a promoted the proliferative potential of gastric adenocarcinoma cell line MGC803. We also identified IL6R as a direct target gene for miR-23a using a fluorescent reporter assay. The mRNA and protein levels of IL6R were both inversely correlated with the miR-23a expression level. Our results demonstrate that miR-23a can target IL6R and promote the growth activity of gastric adenocarcinoma cells in vitro. The downregulation of IL6R by miR-23a may explain why the suppression of miR-23a can inhibit gastric cancer cell proliferation.
FEBS Journal 09/2010; 277(18):3726-34. · 3.79 Impact Factor
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ABSTRACT: MicroRNAs (miRNAs) are emerging as a class of small regulated RNAs, and the alterations of miRNAs are implicated in the initiation and progression of human cancers. Our study shows that inhibition of miR-20a in OVCAR3 ovarian cancer cell line could suppress, whereas overexpression of miR-20a could enhance cell long-term proliferation and invasion. We also confirmed amyloid precursor protein (APP) as a direct target gene of miR- 20a. Furthermore, suppression of APP expression could also promote ovarian cancer cell proliferation and invasion, which is consistent with the results of miR-20a overexpression. Therefore, we concluded that the regulation of APP is an important mechanism for miR-20a to promote proliferation and invasion in ovarian cancer cells.
Acta Biochimica et Biophysica Sinica 05/2010; 42(5):318-24. · 1.38 Impact Factor
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ABSTRACT: Liver targeted micelles were successfully constructed via self-assembly of glycyrrhetinic acid (GA)-modified poly(ethylene glycol)-b-poly(gamma-benzyl l-glutamate) (GA-PEG-PBLG) block co-polymers, which were fabricated via ring opening polymerization of gamma-benzyl l-glutamate N-carboxyanhydride monomer initiated by GA-modified PEG. The in vivo biodistribution and the in vitro cellular uptake of these micelles were investigated. The results showed that the relative uptake of doxorubicin (DOX)-loaded micelles (DOX/GA-PEG-PBLG) in liver was much higher than in other tissues, and the resulting DOX concentration in liver was about 2.2-fold higher than that from the micelles without modification by GA. Moreover, the cellular uptake study demonstrated that the introduction of GA to the micelles could significantly increase the affinity for human hepatic carcinoma 7703 cells, which induced a 3.7-fold higher endocytosis than unmodified ones. The cytotoxicity of DOX/GA-PEG-PBLG micelles (IC(50) 47 ngml(-1)) was much higher than that of free DOX (IC(50) 90 ngml(-1)). These results indicate that GA-modified micelles have great potential in liver targeting therapy.
Acta biomaterialia 05/2010; 6(10):3927-35. · 3.98 Impact Factor
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ABSTRACT: To express and purify the fusion protein of TPT1 (tumor protein translationally-controlled 1) in prokaryocytes and to prepare rabbit anti-TPT1 antibody.
The expression vector pRSETA(2)-TPT1 was reconstructed and transformed into BL21 (DE3). TPT1 fusion protein was induced by IPTG and the TPT1 fusion protein purified by Ni-NTA His Bind resin was used to immunize the rabbit. The titer of the polyclonal antibody was detected by ELISA and its specificity was analyzed by Western blot and IF (immunofluorescence).
The TPT1 fusion protein was highly expressed in E.coli and specific polyclonal antibody was obtained after the immunization.
The recombinant prokaryotic expression vector pRSETA(2)-TPT1 is successfully constructed and high expression of TPT1 is induced in E.coli, which may lay the basis for further study of the development and treatment of tumors and other diseases.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 03/2010; 26(3):246-9.
