Effective tools for RNA-derived therapeutics: siRNA interference or miRNA mimicry

Background/Aims: Tonsillectomy may be an important method to achieve a long-term remission of IgAN, but patients’ physical status may limit their access to this surgery. We proposed an encouraging solution through inhibiting GADD34 expression in order to promote tonsillar mononuclear cells (TMCs) apoptosis and reduce nephropathic IgA secretion. Methods: A total of 12 IgAN and 9 non-IgAN patients were involved from March 2015 to May 2016. After TMCs were extracted by density gradient centrifugation and stimulated by inactivated hemolytic streptococcus, the mRNA and protein expression of GADD34, GRP78, CHOP, Bcl-2, Bcl-XL, AID, Iα-Cα, and cleaved caspase-3 were examined by fluorescent RT-PCR and Western blotting. Guanabenz treatment and siRNA interference were applied to downregulate GADD34 in tonsillar mononuclear cells from IgAN patients, and P-eIF2α expression was examined by Western Blotting. Cell apoptosis was evaluated by Annexin V FITC/PI flowcytometry, and IgA secretion in cultural supernatant was inspected by enzyme linked immunosorbent assay. Results: After stimulation, the expression of GADD34 was significantly increased in IgAN patients (P< 0.05). Cell apoptosis was mitigated and IgA secretion level was elevated (P< 0.05). To be noticed, CHOP expression had no significant difference between two groups. After guanabenz treatment and siRNA interference, a prolonged elevation of P-eIF2α expression was observed. Cell apoptosis was reinforced and IgA secretion level was decreased (P< 0.05). Conclusion: GADD34 may be a potential therapeutic target for IgAN treatment due to its effect on cell apoptosis.

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Suppression of vascular endothelial growth factor via siRNA interference modulates the biological behavior of human nasopharyngeal carcinoma cells

Japanese Journal of Radiology ◽

10.1007/s11604-011-0603-9 ◽

2011 ◽

Vol 29 (9) ◽

pp. 615-622 ◽

Cited By ~ 6

Author(s):

Hai B. Zhou ◽

Yi F. Yin ◽

Yan Hu ◽

Xin Li ◽

Li Y. Zou ◽

...

Keyword(s):

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Nasopharyngeal Carcinoma ◽

Carcinoma Cells ◽

Biological Behavior ◽

Vascular Endothelial ◽

Human Nasopharyngeal Carcinoma ◽

Sirna Interference ◽

Endothelial Growth Factor

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Effect of siRNA interference of ubiquitin-specific protease 9X on apoptosis and growth of diffuse large B cell lymphoma cell line

Materials Express ◽

10.1166/mex.2020.1653 ◽

2020 ◽

Vol 10 (3) ◽

pp. 446-453

Author(s):

Wei Peng ◽

Meizuo Zhong ◽

Youhong Tang

Keyword(s):

B Cells ◽

Protein Expression ◽

B Cell ◽

Cell Apoptosis ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Large B Cell Lymphoma ◽

Specific Protease ◽

Sirna Interference ◽

Large B Cell

Ubiquitin-specific protease 9X (USP9X) is crucial in the diagnosis and treatment of many tumor types, but its role in Diffuse Large B Cell Lymphoma (DLBCL) has not been determined. The current study aimed to examine the effects of RNA interference on USP9X expression, and subsequently on the bioactivity of DLBCL Farage and Pfeiffer cells. There were two groups in the study: USP9X-siRNA and NC. USP9X siRNA was transiently transferred into DLBCL cells by Cationic liposome. The total RNA was extracted using Fe2O3 and was retrieved into the DNA using the MagBeads Total RNA Extraction Kit. The protein expression of USP9X in Farage, Pfeiffer, and normal human B cell line at the cellular level was observed by Western blot. The Farage and Pfeiffer cells were infected with USP9X-siRNA. Cell apoptosis and cell growth viability were analyzed by flow cytometry and CCK8, Mcl-1 protein, a potential target of USP9X, and apoptosis factor proteins (such as Bak, Cytochrome C, Caspase 3, Caspase 8, PARP) were detected by Western blot after siRNA interference. The results showed that the protein expression of USP9X in malignant B cells was four times higher than that of the normal B cells. Inhibition of USP9X reduced the Mcl-1 activity, and increased the caspase-3, Bak and Cytochrome C activity. In the malignant B cells, Mcl-1 and Bak were binding in vivo; Bak was a new partner of Mcl-1. Inhibition of USP9X reduced cell proliferation and increased apoptosis. The expression of USP9X is upregulated in Diffuse large B cell lymphoma cells, Farage, and Pfeiffer. Inhibition expression of USP9X may induce cell apoptosis, inhibit cell growth, and downregulate Mcl-1 protein expression in Diffuse large B cell lymphoma cells, Farage, and Pfeiffer. USP9X has the ability in regulating cell apoptosis.

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Effect of Glucose Metabolism Related Protein-1 on Proliferation and Apoptosis of MIN6 Cells

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2020.2229 ◽

2020 ◽

Vol 10 (2) ◽

pp. 218-222

Author(s):

Yugui Zhang ◽

Yueya Kang ◽

Lixia Zheng ◽

Zhugui Zheng ◽

Jinshui He

Keyword(s):

Cell Proliferation ◽

Glucose Metabolism ◽

Related Protein ◽

Min6 Cells ◽

Min6 Cell ◽

Proliferation And Apoptosis ◽

Sirna Interference ◽

Proliferation Ability ◽

Effect Of Glucose

Apoptosis of islet β cells participates in type 1 diabetes. Exendin-4 promotes islet β cell proliferation and inhibits apoptosis. Glucose metabolism related protein-1 (GMRP-1) has the similar effects on islet β cell. Whether the effect of exendin-4 islet β-cells is regulated by GMRP-1 is not clear. GMRP-1 siRNA or pEGFP-C3-GMRP-1 recombinant plasmid was transfected into MIN6 cells and treated with t-BHP in the presence of exendin-4 (0, 50, 100 and 200 nmol/L) followed by analysis of cell proliferation ability by CCK8 assay, apoptosis by flow cytometry, as well as GMRP-1 protein level by Western blot. After transfection of the plasmid with high expression of GMRP-1, the number of MIN6 cells was increased significantly, and the apoptosis rate was reduced (P < 0.05). With increased exendin-4 concentration, the number of cell proliferation was increased gradually with reduced cell apoptosis (P < 0.05). After the expression of GMRP-1 was decreased after GMRP-1 siRNA interference fragment transfection, the number of MIN6 cells was decreased significantly and the apoptosis was elevated (P < 0.05), whereas, exendin-4’s effect on the proliferation of MIN6 cells and inhibition of apoptosis was attenuated. GMRP-1 affects the proliferation and apoptosis of MIN6 cells, and regulates exonin-4-induced MIN6 cell proliferation and apoptosis.

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Effect of siRNA interference on nerve growth factor in intervertebral disc inflammation rats

Asian Pacific Journal of Tropical Medicine ◽

10.1016/s1995-7645(14)60127-9 ◽

2014 ◽

Vol 7 (9) ◽

pp. 739-743

Author(s):

Ming-Lei Lang ◽

Ai-Lin Qin ◽

Jian-Min Li ◽

Peng Fu

Keyword(s):

Nerve Growth Factor ◽

Growth Factor ◽

Intervertebral Disc ◽

Nerve Growth ◽

Sirna Interference

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Lentivirus-mediated siRNA interference targeting SGO-1 inhibits human NSCLC cell growth

Tumor Biology ◽

10.1007/s13277-011-0284-0 ◽

2011 ◽

Vol 33 (2) ◽

pp. 515-521 ◽

Cited By ~ 11

Author(s):

Lili Liu ◽

Ning Zhang ◽

Jiayu Liu ◽

Jie Min ◽

Ningqiang Ma ◽

...

Keyword(s):

Cell Growth ◽

Nsclc Cell ◽

Sirna Interference ◽

Human Nsclc

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TLR4 Signaling in MPP+-Induced Activation of BV-2 Cells

Neural Plasticity ◽

10.1155/2016/5076740 ◽

2016 ◽

Vol 2016 ◽

pp. 1-9 ◽

Cited By ~ 19

Author(s):

Peng Zhou ◽

Ruihui Weng ◽

Zhaoyu Chen ◽

Rui Wang ◽

Jing Zou ◽

...

Keyword(s):

Western Blot ◽

Mtt Assay ◽

Inflammatory Responses ◽

Cell Model ◽

Critical Role ◽

Control Group ◽

Immunofluorescence Technique ◽

Rt Pcr ◽

Sirna Interference

Aims.This work was conducted to establish anin vitroParkinson’s disease (PD) model by exposing BV-2 cells to 1-methyl-4-phenylpyridinium (MPP+) and exploring the roles of TLR2/TLR4/TLR9 in inflammatory responses to MPP+.Methods/Results.MTT assay showed that cell viability of BV-2 cells was 84.78 ± 0.86% and 81.18 ± 0.99% of the control after incubation with 0.1 mM MPP+for 12 hours and 24 hours, respectively. Viability was not significantly different from the control group. With immunofluorescence technique, we found that MPP+incubation at 0.1 mM for 12 hours was the best condition to activate BV-2 cells. In this condition, the levels of TNF-α, IL-1β, and iNOS protein were statistically increased compared to the control according to ELISA tests. Real time RT-PCR and western blot measurements showed thatTLR4was statistically increased after 0.1 mM MPP+incubation for 12 hours. Furthermore, after siRNA interference ofTLR4mRNA, NF-κB activation and the levels of TNF-α, IL-1β, and iNOS were all statistically decreased in this cell model.Conclusion.MPP+incubation at the concentration of 0.1 mM for 12 hours is the best condition to activate BV-2 cells for mimicking PD inflammation in BV-2 cells. TLR4 signalling plays a critical role in the activation of BV-2 cells and the induction of inflammation in this cell model.

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