Alternative splicing of pre-mRNA is an important mechanism for regulating gene function at the post-transcription level and for producing proteomic diversity in higher eukaryotes. The alternative splicing is regulated by the interaction between diverse cis-acting elements and trans-acting factors. Alternative splicing events of oncogenes, tumor suppressor genes and metastasis suppressor genes are associated with the initiation and development of human neoplasms. The protein isoforms sourced from alternative splicing take part in regulating the gene transcription, cell cycle, apoptosis of cells, and playing a role in tumor growth. It is possible for molecular therapy to target directly isoforms of protein produced by alternative splicing or to interfere with the process of alternative splicing.
Alternative Splicing ; genetics ; Humans ; Neoplasms ; genetics ; RNA Precursors ; metabolism ; RNA, Neoplasm ; analysis ; Transcription, Genetic

Objective To investigate the effects of microRNA-18a (miR-18a) on migration and invasion of hepatocellular carcinoma (HCC) cells, and its possible mechanism associated with Dicer l.Methods HepG2 and HepG2.2.15 cells were transfected with miR-18a inhibitor using Lipofectamine. Cell invasion was evaluated by transwell invasion assay, and cell migration was detected by transwell migration and wound-healing assays. Moreover, luciferase reporter assay was used to identify whether Dicer expression was regulated by miR-18a. Real-time RT-PCR and western blot were performed to analyze Dicer 1 expression. In addition, a functional restoration assay was performed to investigate whether miR-18a promotes HCC cell migration and invasion by directly targeting Dicer 1.Results miR-18a inhibitor can suppress the migration and invasion of HCC cells. Furthermore, suppression of Dicer l expression by small interfering RNA essentially abolished the inhibition of cell migration and invasion induced by miR-18a inhibitor, restorating these activities to levels similar to the parental HCC cells. Interestingly, suppression of miR-18a in HCC cells resulted in enhanced expression of Dicer l. In addition, the results of a luciferase assay demonstrated targeted regulation of Dicer l by miR-18a.Conclusion Our findings suggest that miR-18a promotes migration and invasion of HCC cells by inhibiting Dicer l expression.
Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Movement ; DEAD-box RNA Helicases ; genetics ; metabolism ; Hep G2 Cells ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; MicroRNAs ; genetics ; metabolism ; Neoplasm Invasiveness ; Neoplasm Proteins ; genetics ; metabolism ; RNA, Neoplasm ; genetics ; metabolism ; Ribonuclease III ; genetics ; metabolism

OBJECTIVETo study the expression of prostate stem cell antigen (PSCA) at protein and mRNA levels in invasive micropapillary carcinoma of the breast (IMPC) and to analyze the relationship between PSCA expression and clinicopathologic features.

METHODSThe expression of PSCA protein was analyzed by immunohistochemistry (LSAB) in 66 cases of IMPC and 67 cases of invasive ductal carcinoma, not otherwise specified (IDC-NOS). The association between PSCA expression and clinicopathologic features was also analyzed in IMPC. Furthermore, RT-PCR was used to detect PSCA mRNA in 10 cases of primary IMPC and 10 cases of primary IDC-NOS with paired normal breast tissues, each from the same subject.

RESULTSImmunohistochemical analysis revealed the overexpression of PSCA in 47 of 66 (71.2%) cases of IMPC and 35 of 67 (52.2%) IDC-NOS. Statistical analysis showed a significant difference of PSCA expression between IMPC and IDC-NOS (P = 0.024). In IMPC, the expression of PSCA was correlated with lymph nodes metastasis (P = 0.039). RT-PCR showed the mRNA level of PSCA was significantly higher in primary IMPC and IDC-NOS tissue than that in paired normal breast tissue (7/10 and 5/10, respectively), and it was also significantly higher in primary IMPC tissue than that in IDC-NOS tissue.

CONCLUSIONPSCA might play an important role in lymph node metastasis in IMPC.

Antigens, Neoplasm ; genetics ; metabolism ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; genetics ; metabolism ; pathology ; Carcinoma, Papillary ; genetics ; metabolism ; pathology ; Female ; GPI-Linked Proteins ; genetics ; metabolism ; Humans ; Lymphatic Metastasis ; Neoplasm Invasiveness ; Neoplasm Proteins ; genetics ; metabolism ; Neoplasm Staging ; RNA, Messenger ; metabolism

OBJECTIVETo detect the expression of eukaryotic translation initiation factor 5A2(EIF5A2) in pancreatic adenocarcinoma and its correlation with the clinicopathological characteristics and prognosis.

METHODSA total of 73 patients who were treated in our hospital from March 2007 to December 2008 were enrolled in this study. The expression of EIF5A2 in the surgical samples was detected using immunohistochemical staining. Complete clinicopathological data were obtained from all the patients. The potential correlation between EIF5A2 expression and the clinicopathological features, particularly its role in prognosis, were analyzed.

RESULTSOf these 73 patients, 43 had a high EIF5A2 expression. EIF5A2 expression was significantly correlated with the pathological T stage(P<0.001), N stage(P=0.004), M stage(P=0.039), and TNM stage(P=0.005). Kaplan-Meier method demonstrated that the survival was significantly longer in the low EIF5A2 expression group than in the high EIF5A2 expression group(P=0.003). Cox's hazard model showed EIF5A2 was a significant predictor of overall survival in patients with pancreatic adenocarcinoma.

CONCLUSIONEIF5A2 may be a potential predictor of the poor prognosis in patients with pancreatic adenocarcinoma.

Adenocarcinoma ; diagnosis ; metabolism ; Humans ; Neoplasm Staging ; Pancreatic Neoplasms ; diagnosis ; metabolism ; Peptide Initiation Factors ; metabolism ; Prognosis ; RNA-Binding Proteins ; metabolism

OBJECTIVETo explore the association of FMNL2 expression with the metastatic potential of colorectal cancer cells.

METHODSFMNL2 mRNA and protein expressions in 6 human colorectal cancer cell lines were detected by real-time RT-PCR and immunohistochemical method, respectively, and analyzed for their correlations to the in vitro invasiveness of the cell lines evaluated by Boyden assay. In SW620 and SW480/M5 cell lines, the expression of FMNL2 was repressed by FMNL2 short hairpin RNA (shRNA), and the changes in the invasiveness of the cells were observed.

RESULTSFMNL2 was highly expressed in SW480/M5, LoVo and SW620 cells derived from metastatic colorectal cancers in comparison with that in LS174T, SW480 and HT29 cells, which were derived from primary colorectal cancers. In vitro analysis of the cell invasiveness demonstrated that SW480/M5, LoVo and SW620 cells had higher invasiveness than LS174T, SW480 and HT29 in vitro. In SW480/M5 and SW620 cells, transfection with FMNL2 shRNA resulted in significantly lowered cell invasiveness.

CONCLUSIONFMNL2 may play an important role in the invasion and metastasis of colorectal cancer.

Colorectal Neoplasms ; metabolism ; pathology ; Humans ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Proteins ; genetics ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Tumor Cells, Cultured

A trans-splicing ribozyme which can specifically reprogram human telomerase reverse transcriptase (hTERT) RNA was previously suggested as a useful agent for tumor-targeted gene therapy. In this study, we evaluated in vivo function of the hTERT-targeting trans-splicing ribozymes by employing the molecular analysis of expression level of genes affected by the ribozyme delivery into peritoneal carcinomatosis mice model. To this effect, we constructed adenoviral vector encoding the specific ribozyme. Noticeably, more than four-fold reduction in the level of hTERT RNA was observed in tumor nodules by the systemic infection of the ribozyme-encoding virus. Such hTERT RNA knockdown in vivo induced changes in the global gene expression profile, including the suppression of specific genes associated with anti-apoptosis including bcl2, and genes for angiogenesis and metastasis. In addition, specific trans-splicing reaction with the targeted hTERT RNA took place in the tumors established as peritoneal carcinomatosis in mice by systemic delivery of the ribozyme. In conclusion, this study demonstrates that an hTERT-specific RNA replacement approach using trans-splicing ribozyme represents a potential modality to treat cancer.
Animals ; Cell Line ; Gene Expression/*physiology ; Genetic Vectors ; Humans ; Mice ; Neoplasm Metastasis ; Neoplasms/genetics/pathology ; RNA, Catalytic/genetics/*metabolism ; RNA, Messenger/genetics/metabolism ; RNA, Neoplasm/genetics/metabolism ; Telomerase/antagonists & inhibitors/genetics/*metabolism ; Trans-Splicing/*genetics

OBJECTIVE: To investigate the association between the expression of caveolin-1(CAV-1) and the invasion of choriocarcinoma, and to explore the effect of CAV-1 small interfering RNA(siRNA) on the invasion of choriocarcinoma cell line JEG-3. METHODS: (1) Matrigel invasion assay and 3-(4,4)-dimethylthiahiazo (-z-yl)-3,5-di-phenytetrazoliumormide (MTT) assay were used to examine the difference in invasion and proliferation ability between JEG-3 cells and JAR cells;(2) Expression of caveolin-1 gene in the human chorionic villi tissues and chorionicnoma cell lines (JEG-3 cells and JAR cells) were detected by semi-quantitative RT-PCR. (3) The effect of CAV-1 siRNA transfection on the expression of CAV-1 mRNA, and the invasion and proliferation ability of JEG-3 cells were measured by RT-PCR, Matrigel invasion assay and MTT assay. RESULTS: (1) The invasion ability of JEG-3 cell line was stronger than that of JAR cell line (P<0.05), but the difference in proliferation ability between JAR and JEG-3 was not obvious (P>0.05);(2) The expression of caveolin-1 gene in chorionicnoma cell lines was significantly stronger than that in the human normal chorion(P<0.05), and the expression of caveolin-1 gene in JEG-3 cells was stronger than that in the JAR cells (P<0.05). The data suggested that there was significantly positive correlation between caveolin-1 and the invasiveness of chorionicnoma cells (r=0.086,P<0.05);(3) CAV-1 siRNA could knock-out the expression of CAV-1 mRNA, and inhibit the invasion and proliferation ability of chorionicnoma cells. CONCLUSION CAV-1 can promote the invasion ability of chorionicnoma cells. CAV-1 siRNA can inhibit the invasion and proliferation ability of chorionicnoma cells.
Caveolin 1 ; biosynthesis ; genetics ; Choriocarcinoma ; metabolism ; pathology ; Female ; Humans ; Neoplasm Invasiveness ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Tumor Cells, Cultured ; Uterine Neoplasms ; metabolism ; pathology

OBJECTIVE: To investigate the expression and significance of miRNA-324-3p and its target gene WNT2B in tissue specimens of nasopharyngeal carcinoma (NPC) specimens. METHOD: qRT-PCR was used to detect the expression of miRNA-324-3p and WNT2B mRNA, and Western blot was applied to assay the expression of WNT2B protein in 39 cases of NPC specimens and 21 cases of non-carcinoma epithelium. The relationship between their expression levels and clinicopathological characteristics and their correlation with clinical pathological parameters was analyzed. RESULT: The expression of miRNA-324-3p was significantly down-regulated decreased but WNT2B mRNA/protein increased obviously in NPC specimens (P < 0.01). A negative correlation between miRNA-324-3p and WNT2B was spotted (P < 0.05). The expression levels of these markers were closely correlated with T stage, clinic stage and cervical lymph node metastasis (P < 0.05). CONCLUSION The loss of miRNA-324-3p and ectopic WNT2B might co-induce the initiation and progression of NPC.
Carcinoma ; Glycoproteins ; genetics ; metabolism ; Humans ; Lymphatic Metastasis ; MicroRNAs ; metabolism ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; metabolism ; Neoplasm Proteins ; metabolism ; RNA, Messenger ; metabolism ; Wnt Proteins ; genetics ; metabolism

Objective: To establish enzalutamide-resistant human prostate cancer cell lines and screen out the lncRNA and mRNA expression profiles associated with enzalutamide resistance. METHODS: Human prostate cancer cell lines LNCAP and C4-2B were cultured with 10 μmol/L enzalutamide for 6 months in vitro for the establishment of enzalutamide-resistant subclones LNCAP-ENZA and C4-2B-ENZA. The IC50 value and enzalutamide resistance index of each cell line were examined by MTT assay, the expressions of enzalutamide-related genes FL-AR, AR-V7 and HnRNPA1 were determined by Western blot, and the lncRNA and mRNA differential expressions of C4-2B and C4-2B-ENZA were detected by high-throughout lncRNA microarray. RESULTS: Compared with LNCAP and C4-2B, the IC50 values of enzalutamide-resistant subclones LNCAP-ENZA (60.83 μmol/L) and C4-2B-ENZA (88.32 μmol/L) were increased significantly (P < 0.05) and the enzalutamide-resistance indexes of the LNCAP-ENZA and C4-2B-ENZA cells were 4.94 and 4.67, respectively. The expressions of AR-V7 and HnRNPA1 were markedly up-regulated in the LNCAP-ENZA and C4-2B-ENZA cells as compared with those in the LNCAP and C4-2B cells, but that of FL-AR showed no significant change. A total of 1 440 lncRNAs and 1 236 mRNAs were identified as differentially expressed in the C4-2B-ENZA cells. CONCLUSIONS Enzalutamide -resistant human prostate cancer cell subclones LNCAP-ENZA and C4-2B-ENZA were successfully established and enzalutamide resistance-associated lncRNA and mRNA were identified, which may provide some molecular evidence for the management of enzalutamide-resistant human prostate cancer.
Cell Line, Tumor ; drug effects ; Drug Resistance, Neoplasm ; Humans ; Male ; Phenylthiohydantoin ; analogs & derivatives ; pharmacology ; Prostatic Neoplasms ; drug therapy ; genetics ; pathology ; RNA, Long Noncoding ; metabolism ; RNA, Messenger ; metabolism ; RNA, Neoplasm ; metabolism ; Receptors, Androgen

OBJECTIVETo investigate that the role of Axin in regulating the invasion and migration ability of lymphoma cells and explore the molecular mechanisms.

METHODSThe expressions of Axin, β-catenin, MMP7, and MMP9 were detected in different lymphoma cell lines by RT-PCR and Western blotting. A lymphoma cell line with low Axin expressions was transiently transfected with pCMV5-HA-Axin and pcDNA5-His-β-catenin plasmid, and the expressions of β-catenin, MMP7, and MMP9 mRNA and protein were observed. A lymphoma cell model stably overexpressing Axin was transfected with AXIN-shRNA and β-catenin-shRNA, and the changes in β-catenin, MMP7, and MMP9 cexpressions were observed. The changes in the invasion and migration abilities of this cell model were assessed following Axin knockdown.

RESULTSIn the lymphoma cell lines tested, the Axin expression showed a negative correlation with β-catenin, MMP7, and MMP9 expressions. In Raji cells with a low Axin expression, overexpression of Axin resulted in decreased expressions of β-catenin, MMP7, and MMP9 at the protein levels but not the mRNA levels, and overexpression of β-catenin obviously increased MMP7 and MMP9 mRNA and protein expressions. In the cells with stable Axin overexpression, Axin knockdown caused increased expressions of β-catenin, MMP7, and MMP9 at the protein levels but not the mRNA levels, while β-catenin knockdown caused lowered expressions of MMP7 and MMP9 and suppressed cell invasion and migration.

CONCLUSIONIn lymphoma cells, Axin overexpression can decrease the expression of β-catenin, which in turn decreases the expressions of MMP7 and MMP9 to inhibit the cell invasion and migration.

Axin Protein ; genetics ; metabolism ; Cell Line, Tumor ; Down-Regulation ; Gene Knockdown Techniques ; Humans ; Lymphoma ; genetics ; metabolism ; pathology ; Matrix Metalloproteinase 7 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Neoplasm Invasiveness ; Neoplasm Metastasis ; RNA, Messenger ; RNA, Small Interfering ; Transfection ; beta Catenin ; metabolism