RNA therapeutics inhibit the expression of specific proteins/RNAs by targeting complementary sequences of corresponding genes or encode proteins for the synthesis desired genes to treat genetic diseases. RNA-based therapeutics are categorized as oligonucleotide drugs (antisense oligonucleotides, small interfering RNA, RNA aptamers), and mRNA drugs. The antisense oligonucleotides and small interfering RNA for treatment of genetic diseases have been approved by the FDA in the United States, while RNA aptamers and mRNA drugs are still in clinical trials. Chemical modifications can be applied to RNA drugs, such as pseudouridine modification of mRNA, to reduce immunogenicity and improve the efficacy. The secure and effective delivery systems such as lipid-based nanoparticles, extracellular vesicles, and virus-like particles are under development to address stability, specificity, and safety issues of RNA drugs. This article provides an overview of the specific molecular mechanisms of eleven RNA drugs currently used for treating genetic diseases, and discusses the research progress of chemical modifications and delivery systems of RNA drugs.
Aptamers, Nucleotide ; RNA, Small Interfering/therapeutic use* ; RNA, Messenger ; Oligonucleotides, Antisense/therapeutic use*

MicroRNAs (miRNAs) are a class of highly abundant non-coding RNA molecules that are involved in several biological processes. Many miRNAs are often deregulated in several diseases including cancer. There is substantial interest in exploiting miRNAs for therapeutic applications. In this editorial, we briefly review current advances in the use of miRNAs or antisense oligonucleotides (antagomirs) for such therapies. One of the key issues related to therapy using miRNAs is degradation of naked particles in vivo. To overcome this limitation, delivery systems for miRNA-based therapeutic agents have been developed, which hold tremendous potential for improving therapeutic outcome of cancer patients.
Drug Delivery Systems ; methods ; Gene Expression Regulation, Neoplastic ; Genetic Therapy ; Humans ; MicroRNAs ; genetics ; metabolism ; therapeutic use ; Neoplasms ; genetics ; metabolism ; therapy ; Oligonucleotides, Antisense ; therapeutic use

BACKGROUNDTelomerase activity is found in 85%-90% of all human cancers but not in their adjacent normal cells. Human telomerase reverse transcriptase (hTERT) is an essential component in the telomerase complex that plays an important role in telomerase activity. This study investigated the effect of the telomerase inhibition with an hTERT antisense oligodeoxynucleotide (ODN) in bladder cancer cells (T24) on tumor necrosis factor-alpha (TNF-alpha)-induced apoptosis.

METHODSAntisense phosphorothioate oligodeoxynucleotide (AS PS-ODN) was synthesized and purified. Telomerase activity was measured by polymerase chain reaction enzyme-linked immunoassay (PCR-ELISA). hTERT mRNA expression was measured by reverse transcription polymerase chain reaction (RT-PCR) assay and a gel-image system. hTERT protein was detected by immunochemistry and flow cytometry. Cell viability was measured by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium (MTT) assay. Cell apoptosis was observed by a morphological method and determined by flow cytometry.

RESULTSAS PS-ODN significantly inhibited telomerase activity and decreased the levels of hTERT mRNA which preceded the decline in the telomerase activity. AS PS-ODN significantly reduced the percentage of positive cells expressing hTERT protein following the decline of hTERT mRNA levels. There was no difference seen in the telomerase activity, hTERT mRNA expression or the protein levels between the sense phosphorothioate oligodeoxynucleotide (SPS-ODN) and the control group. AS PS-ODN treatment significantly decreased the cell viability and enhanced the apoptotic rate of T24 cells in response to TNF-alpha while there was no difference in cell viability and apoptotic rate between the S PS-ODN and the control group.

CONCLUSIONSAS PS-ODN can significantly inhibit telomerase activity by downregulating the hTERT mRNA and protein expression. Treatment with AS PS-ODN may be a potential and most promising strategy for bladder cancer with telomerase activity.

Apoptosis ; Cell Line, Tumor ; Flow Cytometry ; Humans ; Oligonucleotides, Antisense ; therapeutic use ; RNA, Messenger ; analysis ; Telomerase ; analysis ; antagonists & inhibitors ; genetics ; Thionucleotides ; therapeutic use ; Tumor Necrosis Factor-alpha ; physiology ; Urinary Bladder Neoplasms ; enzymology ; pathology ; therapy

Cell Line, Tumor ; Genes, ras ; Humans ; Oligonucleotides, Antisense ; therapeutic use ; Pancreatic Neoplasms ; genetics ; therapy ; Point Mutation ; RNA, Messenger ; analysis

Antisense oligonucleotides (ASODN) for therapy is a genetic technology which is based on the base-complementary principle. DNA or RNA sequence synthesized by biotechnology is transferred into the target cells to form mRNA-DNA or mRNA-RNA double strand for inhibiting the expression of target genes. In this way we can control and treat some diseases. The development of antisense oligonucleotides drugs has opened a new area of genetic pharmacology. This paper reviews its classifications, mechanics and its wide application in the treatment of viral infection, tumor and cardiovascular diseases. At the same time we pose the problems that need solving.
Arterial Occlusive Diseases ; therapy ; Humans ; Neoplasms ; therapy ; Oligonucleotides, Antisense ; pharmacology ; therapeutic use ; Virus Diseases ; therapy

OBJECTIVETo observe the effect of survivin antisense oligodeoxynucleotide (ASODN) on the apoptosis of human carcinoma of larynx cell line Hep2 and the inhibitory rate in nude mice model so as to discuss the selective blocking activity of antisense technique on gene expression seeking a new way for gene therapy of carcinoma of larynx.

METHODSAntisense oligodeoxynucleotides survivin were transformed into human carcinoma of larynx cell line Hep2 by liposome Lipofectamine 2000. Within 72 h after transfection, 3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) assay was used to detect cellular proliferation. Forty eight hours after transfection, reverse transcription-polymerase chain reaction (RT-PCR) assay was used to observe the expression of survivin gene, Western Blot assay for the protein, and terminal deoxynucleotide mediated nick end labeling (TUNEL) and flow cytometer for cellular apoptosis.

RESULTSCellular inhibition rate of 72 h went up to 52. 5% and 71.4% at 1.0 micromol/L and 2.0 micromol/L value in Lipo-ASODN groups which differed statistically remarkably (P = 0.046), higher than that in controls in MTT assay (P =0. 003 and 0. 0004). Forty eight hours after transfection survivin gene expression in Lipo-ASODN groups were less than that in control group in RT-PCR assay. Survivin protein expression decreased in Western blot. In TUNEL assay, nuclear positive staining was observed and the apoptosis peak was observed in flow cytometer test, which were absent in controls. In nude mice of carcinoma of larynx model, the inhibitory rate in Lipo-ASODN groups got up to 48.1% and 61.3% higher than that of controls (P < 0.004 and 0. 0006), which differed remarkably (P = 0.032) in a dose-dependently way.

CONCLUSIONSThe findings showed that the expression of survivin gene and protein induced cellular apoptosis in Hep2 cells after transfection of Lipo-ASODN and that the carcinoma of larynx in the nude mice model were inhibited by Lipo-ASODN which suggested that antisense technique can be an effective means in the gene therapy of carcinoma of larynx.

Animals ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; Laryngeal Neoplasms ; therapy ; Male ; Mice ; Mice, Nude ; Microtubule-Associated Proteins ; genetics ; pharmacology ; therapeutic use ; Oligonucleotides, Antisense ; genetics ; pharmacology ; therapeutic use ; Transfection

OBJECTIVE: To explore the feasibility of antisense oligodeoxynucleotide (ASODN) targeting survivin gene for cisplatin resistant human lung adeno-carcinoma xenograft in nude mice. METHODS: Cisplatin resistant cell lines A549/CDDP were cultured routinely with RPMI1640 medium. A549/CDDP cells were subcutaneously implanted in nude mice to establish cisplatin resistant xenograft animal models. After survivin ASODN mediated by cytofectin was directly injected into xenograft in 5 places. The volumes and weight of tumor mass were detected, respectively, and then tumor growth inhibitory rate and tumor growth index were calculated. Reverse transcription-polymerase chain reaction (RT-PCR) and immunochemohistology assay were performed to detect the expression level of survivin mRNA and protein. RESULTS: In mice treated with single ASODN, the tumor growth inhibitory rate and tumor growth index was 35.4% and 4.23+/-0.4456. The difference of the tumor growth inhibitory rate and tumor growth index between blank control group and ASODN group was significant (P<0.05). While combined ASODN with cisplatin,the anticancer efficacy was far more significant and the tumor growth inhibitory rate was enhanced to 63.7%. The tumor growth index, however, reduced to 1.700+/-0.436, which was obviously significant,compared with the cisplatin group and other controls (P<0.05). The anticancer efficacy was even more obvious than that of ASODN group (P<0.05). Significant down-regulation of survivin mRNA and protein level expression in tumor tissues of ASODN group and ASODN and cisplatin group was detected by RT-PCR and immunochemohistology assay, respectively (P<0.05). CONCLUSION Survivin ASODN mediated by cytofectin can inhibit the cisplatin resistant tumor growth by direct intra-tumoral injection. The anticancer efficacy may be associated with the down regulation of survivin expression. ASODN targeting survivin gene can be a supportive therapy to cisplatin resistant lung cancer, while the clinical effective values need further exploration.
Adenocarcinoma ; pathology ; therapy ; Animals ; Cisplatin ; pharmacology ; Drug Resistance, Neoplasm ; Female ; Genetic Therapy ; Humans ; Inhibitor of Apoptosis Proteins ; Lung Neoplasms ; pathology ; therapy ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microtubule-Associated Proteins ; genetics ; therapeutic use ; Neoplasm Transplantation ; Oligonucleotides, Antisense ; therapeutic use ; Repressor Proteins ; Survivin

OBJECTIVETo investigate the synergistic effects of antisense HIF-1alpha gene therapy combined with B7-1-mediated immunotherapy on cancer treatment.

METHODSAntisense HIF-1alpha and B7-1 expression vector were constructed. Lymphoma cells EL-4 were injected subcutaneously into C57BL/6 mice and transplanted lymphomas were established. The mice received either antisense HIF-1alpha, B7-1, or a combinational agent, complexed with DOTAP cationic liposomes. The tumor growth in the mice was monitored. Expression of HIF-1alpha, B7-1 and VEGF were detected by immunohistochemistry and Western blotting. The tumor blood vessels were immunostained with CD31- antibodies and the tumor vascular density was assessed by light microscopy.

RESULTSGene transfer of plasmid expressing the encoded antisense HIF-1alpha inhibited VEGF expression and reduced vascular density in the tumors, eradicated tumors in diameter smaller than 0.1 cm and only retarded the growth of larger tumors. Whereas combination of antisense HIF-1alpha gene therapy and B7-1 immunotherapy eradicated all tumors in diameter of 0.4 cm.

CONCLUSIONAntisense HIF-1alpha blocks tumor hypoxia pathway by downregulating VEGF expression, reduction of vascular density and enhances B7-1-mediated immunotherapy. Strategies that target HIF-1 may have therapeutic potential in cancer treatment and are worthy of further studying.

Animals ; B7-1 Antigen ; genetics ; therapeutic use ; Gene Transfer Techniques ; Genetic Therapy ; methods ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; therapeutic use ; Lymphoma ; immunology ; therapy ; Male ; Mice ; Mice, Inbred C57BL ; Neovascularization, Pathologic ; therapy ; Oligonucleotides, Antisense ; genetics ; therapeutic use

OBJECTIVETo define changes in clusterin expression following short-term neoadjuvant hormone therapy (NHT) and its biological significance in prostate cancer tissues.

METHODSTwenty-six archival radical prostatectomy (RP) specimens without receiving NHT, 19 needle biopsies and corresponding 19 RP specimens following 3-month NHT, were subjected to immunohistochemical clusterin staining.

RESULTSStaining for clusterin was mainly found in cytoplasm and part of extracellular matrix. Clusterin expression was significantly greater in RP specimens with preoperative NHT (t = 2.91, P < 0.01); Needle biopsies obtained before NHT consistently demonstrated lower staining intensity (1.42 +/- 0.51) than corresponding RP specimens (2.16 +/- 0.60) following 3-month NHT (t = 7.10, P < 0.01).

CONCLUSIONSUpregulation of clusterin in part accounts for malignant progression of prostate cancer through its anti-apoptotic action following androgen withdrawal. These findings support that adjuvant therapy targeting clusterin may enhance androgen ablation therapy in advanced prostate cancer.

Aged ; Clusterin ; genetics ; metabolism ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoadjuvant Therapy ; methods ; Oligonucleotides, Antisense ; therapeutic use ; Prostatic Neoplasms ; metabolism ; pathology ; therapy

With the proceeding of the human genome program(HGP), new genes related to male reproduction have been cloned continuously by different methods. However, investigation of the gene function is still at the rudimentary stage. In spite of the advances in the common methods of studying gene function, some special methods have yet to be developed concerning the study of the genes related to male reproduction. This paper summarizes the current methods of study on gene function relevant to male reproduction.
Animals ; Gene Targeting ; Gene Transfer, Horizontal ; Genes ; physiology ; Genetic Vectors ; genetics ; Humans ; Male ; Models, Animal ; Oligonucleotides, Antisense ; therapeutic use ; Reproduction ; genetics ; Reverse Transcriptase Polymerase Chain Reaction