ShRNA‐mediated matrix metalloproteinase‐2 gene silencing protects normal cells and sensitizes cancer cells against ionizing‐radiation induced damage

Gugalavath, Shailender; Patanla, Kiranmayi; Malla, Rama Rao

doi:10.1002/jcb.29369

Cited by 8 publications

(3 citation statements)

References 105 publications

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“…Silencing of Ku70 transcription and translation is known to be more significant than the mitigation of Ku70 acetylation modifications, indicating a Ku70 dosage-dependent elevation of Bax in cells (Ding et al 2021;Wang et al 2020). The Bax then leads to the release of pro-apototic factors, such as Cyt-c from the mitochondria and the activation of the caspases, resulting in the initiation of the intrinsic pathway of apoptosis (Shailender et al 2020;Wang et al 2019). Consistently, our data revealed that silencing of Ku70 upregulate the level of Bax.…”

Section: Discussionsupporting

confidence: 84%

MiR-1224 downregulation inhibits OGD/R-induced hippocampal neuron apoptosis through targeting Ku protein

Wan

Xiao

2021

Metab Brain Dis

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Ischemic cerebrovascular disease is the main cause of disability due to stroke. This study aimed to investigate the function of miR-1224 in OGD/R-induced hippocampal neuron apoptosis, as well as the regulatory mechanism of miR-1224 in ischemic cerebrovascular disease. The oxygen-glucose deprivation/reperfusion (OGD/R) model of primary mouse hippocampal neurons was established. RT-qPCR detected miR-1224, Ku70 and Ku86 levels. Western blotting was applied to measure the expression of Ku70/86 and apoptosis related proteins. Flow cytometry was used to assess apoptosis. JC-1 fluorescence was performed to test the mitochondrial membrane potential (MMP) in neurons. The double luciferase reporter assay was performed to investigate the relationship between miR-1224 and Ku70/86. OGD/R induced the apoptosis and mitochondrial injury in neuronal cells, while miR-1224 downregulation or Ku70/86 upregulation reversed this phenomenon. Meanwhile, miR-1224 negatively regulated the expression of Ku70/86 in neuronal cells through directly targeting Ku70/86. Furthermore, knockdown of Ku70/86 significantly reversed the inhibitory effect of miR-1224 silencing on apoptosis and mitochondrial injury in OGD/R-treated neuronal cells. Our findings indicated that miR-1224 downregulation suppressed OGD/R-induced hippocampal neuron apoptosis by targeting Ku protein, suggesting that miR-1224 could serve as a new target for ischemic cerebrovascular disease treatment.

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Section: Discussionsupporting

confidence: 84%

MiR-1224 downregulation inhibits OGD/R-induced hippocampal neuron apoptosis through targeting Ku protein

Wan

Xiao

2021

Metab Brain Dis

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“…The results showed that compared with the non‐irradiation group, the cells in the irradiation group were all blocked in the G2/M phase; Compared with the irradiation control group, 20 and 40 μg/mL tCQA significantly inhibited the effect of 8Gy IR on G2/M phase of blockade (Figure 2D,E). According to reports, G2/M phase is regulated by CDK1‐cyclin B1 complex 23 ; CDK1 activation is a necessary condition for G2 phase to enter M phase; Thr‐161 phosphorylation can activate CDK1 24,25 ; After the complex formed by cyclin B1 and CDK1 is activated, the cell cycle enters M phase from G2 phase; Radiation injury is often accompanied by decreased expression of cyclin B1 25–27 . Western blot showed that 8Gy IR degraded cyclin B1 and inhibited CDK1 activation; however, tCQA could significantly increase the protein levels of cyclin B1 and CDK1 (pThr‐161; Figure 2F).…”

Section: Resultsmentioning

confidence: 99%

3,4,5‐O‐tricaffeoylquinic acid alleviates ionizing radiation‐induced injury in vitro and in vivo through regulating ROS/JNK/p38 signaling

Liu

et al. 2021

Environmental Toxicology

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Ionizing radiation (IR) brings many health problems to humans, causing damage to the digestive system, hematopoietic system, and immune system. Natural compounds derived from plants have attracted widespread attention due to their low toxicity. Here, we found that 3,4,5‐O‐tricaffeoylquinic acid (tCQA) extracted from natural plant Azolla imbricata could significantly alleviate the systemic damage in mice caused by IR. In order to further explore the molecular mechanism of the radioprotective effect of tCQA, in vitro experiments confirmed that tCQA could attenuate the cytotoxic effect of IR on the colonic epithelial cell line NCM460 and alleviate the IR‐induced mitochondrial dysfunction characterized by the decrease of mitochondrial transmembrane potential, ROS production, and caspase‐dependent apoptosis. In addition, the generation of ROS induced by H2O2 could also be reversed by tCQA. Then, Western blot demonstrated that tCQA could reverse the MAPK signaling pathway activated by IR. However, the inhibitory effect of tCQA on JNK and P38 levels activated by the JNK agonist anisomycin is not obvious; meanwhile, tCQA could inhibit the activation of JNK/P38 induced by H2O2, which suggests that tCQA might inhibit the JNK/P38 signaling pathway by reducing ROS. In short, tCQA inhibits the generation of ROS caused by IR, and then regulates the activity of caspase in the mitochondrial pathway by inhibiting the JNK/P38 signaling pathway, thereby alleviating the apoptosis of NCM460. This research provides an experimental basis for the development of new types of radioprotective agents for medical diagnosis and radiotherapy.

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“…RNA interference (RNAi) has become one of the powerful tools in recent times to suppress the expression of speci c gene of interest [17][18], as a therapeutic option in disease management [19][20][21][22], as cancer treatment [23][24][25] and for understanding regulatory function of a gene by dsRNA molecules [26]. Suppression of gene expression achieved by cleavage of dsRNA molecules by ribonuclease protein (Dicer) followed by loading of siRNA molecules into RISC (RNA induced silencing complex) and then, guide strand of the siRNA molecule would guide the RISC towards the target mRNA sequence complementary with the siRNA for cleavage of target mRNA [27][28][29][30][31].…”

Section: Discussionmentioning

confidence: 99%

S ilencing of SCD and SREBP1 Genes by Short Hairpin RNAs in Chicken Hepatic Cells in Vitro

Sagar

Prasad

Kumar

et al. 2022

Preprint

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RNA interference by short hairpin RNAs (shRNAs) is a widely used post transcriptional silencing mechanism for suppressing expression of the target gene. In the current study, five shRNA molecules each against SCD and SREBP1 genes involved in denovo lipid biosynthesis were designed upon considering parameters such as secondary structures of shRNAs, mRNA target regions, GC content and thermodynamic properties (ΔG overall, ΔG duplex and ΔG break-target), synthesized and cloned in pENTR/U6 entry vector to knockdown the expression of SCD and SREBP1 genes. After transfection of these shRNA constructs into the chicken embryonic hepatocytes, expressions of the target genes were monitored by real time PCR. Significant reduction (P<0.05) in the expression of SCD and SREBP1 genes was observed in hepatocytes. The shRNAs against SCD gene showed the knock down efficiency ranged from 20.4% (shRNA5) to 74.2% (shRNA2). In case of SREBP1 gene, the shRNAs showed knock-down efficiency ranging from 26.8% (shRNA4) to 95.85% (shRNA1). The shRNAs against both the genes introduced in chicken hepatocyte cells did not show any significant impact on expression of immune response genes (IFNA and IFNB) in those cells. These results clearly demonstrated the successful down regulation of the expression of SCD and SREBP1 genes by the shRNA molecules against both the target genes under in vitro condition. It is concluded that the shRNA molecules against SCD and SREBP1 genes showed great potential to silence the expression of these genes under in vitro chicken embryonic hepatocyte cells.

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ShRNA‐mediated matrix metalloproteinase‐2 gene silencing protects normal cells and sensitizes cancer cells against ionizing‐radiation induced damage

Cited by 8 publications

References 105 publications

MiR-1224 downregulation inhibits OGD/R-induced hippocampal neuron apoptosis through targeting Ku protein

MiR-1224 downregulation inhibits OGD/R-induced hippocampal neuron apoptosis through targeting Ku protein

3,4,5‐O‐tricaffeoylquinic acid alleviates ionizing radiation‐induced injury in vitro and in vivo through regulating ROS/JNK/p38 signaling

S ilencing of SCD and SREBP1 Genes by Short Hairpin RNAs in Chicken Hepatic Cells in Vitro

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