micRNA (messenger RNA-interfering RNA)

The field of gene therapy has experienced an insurgence of attention for its widespread ability to regulate gene expression by targeting genomic DNA, messenger RNA, microRNA, and short-interfering RNA for treating malignant and non-malignant disorders. Numerous nucleic acid analogs have been developed to target coding or non-coding sequences of the human genome for gene regulation. However, broader clinical applications of nucleic acid analogs have been limited due to their poor cell or organ-specific delivery. To resolve these issues, non-viral vectors based on nanoparticles, liposomes, and polyplexes have been developed to date. This review is centered on non-viral vectors mainly comprising of cationic lipids and polymers for nucleic acid-based delivery for numerous gene therapy-based applications.

Download Full-text

A conserved CCCH-type zinc finger protein regulates mRNA nuclear adenylation and export

The Journal of Cell Biology ◽

10.1083/jcb.200811072 ◽

2009 ◽

Vol 185 (2) ◽

pp. 265-277 ◽

Cited By ~ 30

Author(s):

Jessica A. Hurt ◽

Robert A. Obar ◽

Bo Zhai ◽

Natalie G. Farny ◽

Steven P. Gygi ◽

...

Keyword(s):

Zinc Finger ◽

Nuclear Export ◽

Messenger Rna ◽

Zinc Finger Protein ◽

Liquid Chromatography Mass Spectrometry ◽

Functional Conservation ◽

Finger Protein ◽

Interfering Rna ◽

Processing Steps ◽

Prior Processing

Coupling of messenger RNA (mRNA) nuclear export with prior processing steps aids in the fidelity and efficiency of mRNA transport to the cytoplasm. In this study, we show that the processes of export and polyadenylation are coupled via the Drosophila melanogaster CCCH-type zinc finger protein CG6694/dZC3H3 through both physical and functional interactions. We show that depletion of dZC3H3 from S2R+ cells results in transcript hyperadenylation. Using targeted coimmunoprecipitation and liquid chromatography mass spectrometry (MS)/MS techniques, we characterize interactions of known components of the mRNA nuclear export and polyadenylation machineries with dZC3H3. Furthermore, we demonstrate the functional conservation of this factor, as depletion of its human homologue ZC3H3 by small interfering RNA results in an mRNA export defect in human cells as well. Nuclear polyadenylated (poly(A)) RNA in ZC3H3-depleted cells is sequestered in foci removed from SC35-containing speckles, indicating a shift from the normal subnuclear distribution of poly(A) RNA. Our data suggest a model wherein ZC3H3 interfaces between the polyadenylation machinery, newly poly(A) mRNAs, and factors for transcript export.

Download Full-text

Inhibition of PTEN Gene Expression by Small Interfering RNA on PI3K/Akt/FoxO3a Signaling Pathway in Human Nasopharyngeal Carcinoma

Technology in Cancer Research & Treatment ◽

10.1177/1533033820917959 ◽

2020 ◽

Vol 19 ◽

pp. 153303382091795

Author(s):

Liang Zhong Yao ◽

Yan Li Zhu ◽

Jun Jie Liu

Keyword(s):

Nasopharyngeal Carcinoma ◽

Signaling Pathway ◽

Messenger Rna ◽

Small Interfering Rna ◽

Quantitative Polymerase Chain Reaction ◽

Pten Gene ◽

Control Group ◽

Expression Levels ◽

Human Nasopharyngeal Carcinoma ◽

Interfering Rna

The objective of this article is to study the effect of inhibiting phosphatase and tensin homolog deleted chromatosome 10 gene on phosphoinositide 3-kinase/protein kinase B ( Akt)/Forkhead homeobox O3a signaling pathway in human nasopharyngeal carcinoma HK-1 cells. Nasopharyngeal carcinoma HK-1 cell lines were divided into PTEN gene interference group (siPTEN), nonspecific small interfering RNA group (siNC), empty vector group (Vector), and no transfection control group (Normal). The mRNA and protein expression levels of PTEN, PI3K, p-Akt, and FoxO3a were detected by real-time fluorescence quantitative polymerase chain reaction and Western blot. Immunofluorescence was used to detect the subcellular localization of PTEN, PI3K, p-Akt, and FoxO3a in HK-1 cells. The proliferation of HK-1 cells was detected by MTT assay, and the apoptosis of HK-1 cells was detected by flow cytometry. Compared with the siNC group, the expression levels of PTEN, FoxO3a messenger RNA, and protein in the siPTEN group were significantly decreased ( P < .05), while the expression levels of PI3K, p-Akt messenger RNA, and protein were significantly increased ( P < .05). The growth rate of HK-1 cells in the siPTEN group was significantly higher than the siNC group ( P < .05), while the apoptosis rate was significantly lower than that of the siNC group ( P < .05). Small interfering RNA can inhibit the expression of PTEN in HK-1 cells, and PTEN can participate in the development of NPC by affecting PI3K/Akt/FoxO3a signaling pathway.

Download Full-text

Patient Centricity Driving Formulation Innovation: Improvements in Patient Care Facilitated by Novel Therapeutics and Drug Delivery Technologies

The Annual Review of Pharmacology and Toxicology ◽

10.1146/annurev-pharmtox-052120-093517 ◽

2022 ◽

Vol 62 (1) ◽

pp. 341-363

Author(s):

Susanne Page ◽

Tarik Khan ◽

Peter Kühl ◽

Gregoire Schwach ◽

Kirsten Storch ◽

...

Keyword(s):

Patient Care ◽

Targeted Delivery ◽

Messenger Rna ◽

Medical Information ◽

Digital Platforms ◽

Therapeutic Modalities ◽

Age Appropriate ◽

Patient Groups ◽

Sites Of Action ◽

Interfering Rna

Innovative formulation technologies can play a crucial role in transforming a novel molecule to a medicine that significantly enhances patients’ lives. Improved mechanistic understanding of diseases has inspired researchers to expand the druggable space using new therapeutic modalities such as interfering RNA, protein degraders, and novel formats of monoclonal antibodies. Sophisticated formulation strategies are needed to deliver the drugs to their sites of action and to achieve patient centricity, exemplified by messenger RNA vaccines and oral peptides. Moreover, access to medical information via digital platforms has resulted in better-informed patient groups that are requesting consideration of their needs during drug development. This request is consistent with health authority efforts to upgrade their regulations to advance age-appropriate product development for patients. This review describes formulation innovations contributingto improvements in patient care: convenience of administration, preferred route of administration, reducing dosing burden, and achieving targeted delivery of new modalities.

Download Full-text

Messenger RNA-interfering complementary RNA

The Dictionary of Genomics, Transcriptomics and Proteomics ◽

10.1002/9783527678679.dg07326 ◽

2015 ◽

pp. 1-1

Keyword(s):

Messenger Rna ◽

Rna Interfering

Download Full-text

Effect of NIMA-related kinase 2B on the sensitivity of breast cancer to paclitaxel in vitro and vivo

Tumor Biology ◽

10.1177/1010428317699754 ◽

2017 ◽

Vol 39 (5) ◽

pp. 101042831769975 ◽

Cited By ~ 3

Author(s):

Yahong Wang ◽

Honghong Shen ◽

Quangui Yin ◽

Tongxian Zhang ◽

Ziyu Liu ◽

...

Keyword(s):

Breast Cancer ◽

Nude Mouse ◽

Messenger Rna ◽

Small Interfering Rna ◽

Ductal Carcinoma ◽

Positive Lymph Node ◽

Disease Rates ◽

Interfering Rna ◽

Inhibit Cell Proliferation

NIMA-related kinase 2B has been known to be an important centrosome regulatory factor. The aim of this study was to investigate the effect of NIMA-related kinase 2B on the sensitivity of breast cancer to paclitaxel. We detected the expression of NIMA-related kinase 2B messenger RNA in MCF-10 cells, including MCF-10A, MCF-10AT, MCF-10DCIS.com , and MCF-10CA1a. The influence of NIMA-related kinase 2B in nude mouse was also detected. The association between NIMA-related kinase 2B and clinicopathological factors was explored in invasive ductal carcinoma tissues. NIMA-related kinase 2B was lowly expressed in the precancerous cells, MCF-10A and MCF-10AT, and it was highly expressed in carcinomatous cells, MCF-10DCIS.com and MCF-10CA1a. The upregulation of NIMA-related kinase 2B can introduce the growth of MCF-10AT cells, knockdown of NIMA-related kinase 2B could remarkably inhibit cell proliferation in MCF-10DCIS.com and MCF-10 CA1a cells. Comparing the volume of the xenografts in nude mouse, we found that the tumors treated by NIMA-related kinase 2B small interfering RNA associated with paclitaxel were the smallest among all the groups. Expression of NIMA-related kinase 2B messenger RNA was associated with higher histological grades, positive lymph node, and high Ki67 index (>20%). The partial response rates were 75.0% in NIMA-related kinase 2B negative (NIMA-related kinase 2B−) patients and 15.8% in NIMA-related kinase 2B++ patients. The progressive disease rates were 10.0% in NIMA-related kinase 2B− patients and 52.6% in NIMA-related kinase 2B++ patients ( p = 0.002). Our findings suggested that NIMA-related kinase 2B could play a role in the development and progression of breast cancer. Combination treatment using NIMA-related kinase 2B small interfering RNA and paclitaxel might be a novel potential therapy method for breast cancer.

Download Full-text

IQGAP1 Is Involved in Enhanced Aggressive Behavior of Epithelial Ovarian Cancer Stem Cell-Like Cells During Differentiation

International Journal of Gynecological Cancer ◽

10.1097/igc.0000000000000394 ◽

2015 ◽

Vol 25 (4) ◽

pp. 559-565 ◽

Cited By ~ 6

Author(s):

Lu Huang ◽

Shanshan Xu ◽

Dongxiao Hu ◽

Weiguo Lu ◽

Xing Xie ◽

...

Keyword(s):

Ovarian Cancer ◽

Stem Cells ◽

Stem Cell ◽

Cancer Stem Cells ◽

Cancer Stem Cell ◽

Messenger Rna ◽

De Novo ◽

Invasion And Metastasis ◽

Spheroid Culture ◽

Interfering Rna

BackgroundWide metastasis is one of characteristics of ovarian cancer. Cancer stem cells, as a source in cancer invasion and metastasis, possess powerful potential of differentiation. Scaffolding IQ domain GTPase-activating protein 1 (IQGAP1) plays a key role in the invasion and metastasis of cancer cells, but IQGAP1’s role in cancer stem cells including ovarian cancer was unclear.MethodsSpheroid culture with serum-free medium was used for enriching ovarian cancer stem cell-like cells (CSC-LCs) from 3AO cell line, and a medium with 10% fetal bovine serum was used to induce the differentiation of CSC-LCs. Immunofluorescence was for detecting the stem markers OCT4 and SOX2. The quantitative real-time-polymerase chain reaction and Western blotting were performed to determine the messenger RNA and protein expression of IQGAP1, respectively. The capacity of cell invasion was evaluated by transwell chamber assay.ResultsOvarian CSC-LCs obtained through spheroid culture showed irregularly elongated appearance, CD24 negative, and OCT4 and SOX2 positive. IQGAP1 expression was decreased in ovarian CSC-LCs compared with parental 3AO cells, but increased de novo during the differentiation of CSC-LCs. Knockdown of IQGAP1 by specific small interfering RNA remarkably weakened invasion capacity of 2-day differentiated ovarian CSC-LCs.ConclusionsIncreased IQGAP1 expression during the differentiation of CSC-LCs is involved in an aggressive cell behavior, which may contribute to metastasis of ovarian cancer.

Download Full-text

Golgi Oncoprotein GOLPH3 Gene Expression Is Regulated by Functional E2F and CREB/ATF Promoter Elements

Genes ◽

10.3390/genes10030247 ◽

2019 ◽

Vol 10 (3) ◽

pp. 247 ◽

Cited By ~ 1

Author(s):

Beatriz Peñalver-González ◽

Jon Vallejo-Rodríguez ◽

Gartze Mentxaka ◽

Asier Fullaondo ◽

Ainhoa Iglesias-Ara ◽

...

Keyword(s):

Cell Cycle ◽

Messenger Rna ◽

Cell Cycle Regulation ◽

Luciferase Reporter ◽

Luciferase Reporter Construct ◽

Daughter Cells ◽

Genes Encoding ◽

E2f Transcription Factors ◽

Cycle Regulation ◽

Interfering Rna

The Golgi organelle duplicates its protein and lipid content to segregate evenly between two daughter cells after mitosis. However, how Golgi biogenesis is regulated during interphase remains largely unknown. Here we show that messenger RNA (mRNA) expression of GOLPH3 and GOLGA2, two genes encoding Golgi proteins, is induced specifically in G1 phase, suggesting a link between cell cycle regulation and Golgi growth. We have examined the role of E2F transcription factors, critical regulators of G1 to S progression of the cell cycle, in the expression of Golgi proteins during interphase. We show that promoter activity for GOLPH3, a Golgi protein that is also oncogenic, is induced by E2F1-3 and repressed by E2F7. Mutation of the E2F motifs present in the GOLPH3 promoter region abrogates E2F1-mediated induction of a GOLPH3 luciferase reporter construct. Furthermore, we identify a critical CREB/ATF element in the GOLPH3 promoter that is required for its steady state and ATF2-induced expression. Interestingly, depletion of GOLPH3 with small interfering RNA (siRNA) delays the G1 to S transition in synchronized U2OS cells. Taken together, our results reveal a link between cell cycle regulation and Golgi function, and suggest that E2F-mediated regulation of Golgi genes is required for the timely progression of the cell cycle.

Download Full-text

Inhibitory effect ofHuRgene small interfering RNA segment on laryngeal carcinoma Hep-2 cell growth

The Journal of Laryngology & Otology ◽

10.1017/s0022215110001015 ◽

2010 ◽

Vol 124 (11) ◽

pp. 1183-1189 ◽

Cited By ~ 5

Author(s):

Z Shen ◽

D Ye ◽

X Zhang ◽

Z Jiang ◽

B Xiao ◽

...

Keyword(s):

Cell Proliferation ◽

Cell Growth ◽

Laryngeal Carcinoma ◽

Messenger Rna ◽

Small Interfering Rna ◽

Tertiary Care ◽

Cyclooxygenase 2 ◽

Dependent Manner ◽

Tertiary Care Centre ◽

Interfering Rna

AbstractObjectives:To investigate the effect of theHuRgene on laryngeal carcinoma Hep-2 cell growth, and to analyse correlations between theHuR, cyclooxygenase-2 and survivin genes.Study design:Experiment study.Setting:Department of Otolaryngology-Head and Neck Surgery, Lihuili Hospital of Ningbo University, Ningbo, a tertiary care centre in China.Methods:Copies of a small interfering RNA segment directed against theHuRgene were transfected into Hep-2 cells using LipofectamineTM2000. The effect of the small interfering RNA segment on Hep-2 cell proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Changes in the expression of theHuR, cyclooxygenase-2 and survivin genes were detected by semi-quantitative reverse transcription polymerase chain reaction analysis. Concentrations of the HuR, cyclooxygenase-2 and survivin proteins were evaluated using Western blotting.Results:Expression of theHuR, cyclooxygenase-2 and survivin genes, as indicated by messenger RNA and protein levels, was suppressed by theHuRgene small interfering RNA segment in a dose-dependent manner. The proliferation indices of all treated groups were significanlty lower than those of control groups (p < 0.05).Conclusions:Impairment ofHuRgene expression, using interfering RNA technology, can significantly suppress Hep-2 cell proliferation and induce apoptosis. TheHuRgene may be an effective target for gene therapy in patients with laryngeal carcinoma.

Download Full-text

Effect of SOCS1 silencing on proliferation and apoptosis of melanoma cells: An in vivo and in vitro study

Tumor Biology ◽

10.1177/1010428317694315 ◽

2017 ◽

Vol 39 (5) ◽

pp. 101042831769431

Author(s):

Sheng-Jia Yu ◽

Zi-Wen Long

Keyword(s):

Messenger Rna ◽

Small Interfering Rna ◽

Melanoma Cells ◽

Janus Kinase ◽

Extracellular Signal ◽

Proliferation And Apoptosis ◽

Protein Expressions ◽

Interfering Rna

This study aimed to investigate the effect of SOCS1 silencing on the proliferation and apoptosis of melanoma cells by in vivo and in vitro studies. Immunohistochemical staining was used to detect SOCS1 expression in melanoma tissues and pigmented nevi. Quantitative real-time polymerase chain reaction and western blotting were applied to detect the messenger RNA and protein expressions of SOCS1 in primary human melanocytes and malignant melanoma cell lines (A375, SK-MEL-5, M14, and MV3). Melanoma cells were assigned into mock, negative small interfering RNA, and SOCS1-small interfering RNA groups. The proliferation, cell cycle and apoptosis, and messenger RNA expression of SOCS1 in MV3 and A375 cells were detected using MTT assay, flow cytometry, and quantitative real-time polymerase chain reaction, respectively. The expressions of SOCS1 protein, extracellular signal–regulated kinase, and janus kinase signal transduction and activators of transcription signaling pathways–related proteins were detected using western blotting. After the establishment of subcutaneous xenograft tumor models in nude mice, the latent period, size, volume and growth speed of xenograft tumors in the mock, negative small interfering RNA, and SOCS1-small interfering RNA groups were examined and compared. The results indicated that positive expression rate of SOCS1 was higher in malignant melanoma tissues than in pigmented nevi. MV3 cells had the highest messenger RNA and protein expressions of SOCS1, followed by A357 cells. Compared with the mock and negative small interfering RNA groups, SOCS1-small interfering RNA group showed lower cell viability, elevated cell apoptosis, more cells in G0/G1 phase and less cells in S and G2/M phases, and decreased messenger RNA and protein expressions of SOCS1, p-ERK1/2, p-JAK2, p-STAT1, and p-STAT3. Compared with the mock and negative small interfering RNA groups, the SOCS1-small interfering RNA group showed longer latent period of tumor, smaller tumor size and volume, and smoother tumor growth curve. To conclude, SOCS1 silencing can inhibit proliferation and induce apoptosis of MV3 and A357 melanoma cells in vivo and in vitro by inhibiting extracellular signal–regulated kinase and janus kinase signal transduction and activators of transcription signaling pathways.

Download Full-text