Modeling qRT-PCR dynamics with application to cancer biomarker quantification

Chervoneva, Inna; Freydin, Boris; Hyslop, Terry

doi:10.1177/0962280216683204

Cited by 8 publications

(5 citation statements)

References 34 publications

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“…cDNA was synthesized using Hifair ® II 1st Strand cDNA Synthesis Kit (YeaSen Biotech, Shanghai, China). Fluorescence qRT-PCR reactions were performed using SYBR Premix Ex Taq Kit (TaKaRa, Dalian, China) (14). Primers used for qRT-PCR are listed in Table 1.…”

Section: Quantitative Real-time Pcr (Qrt-pcr) and Enzymelinked Immunomentioning

confidence: 99%

Toll-like receptor 4 (TLR4) deficiency aggravates dextran sulfate sodium (DSS)-induced intestinal injury by down-regulating IL6, CCL2 and CSF3

Yunjie¹,

Hu²,

Zhao³

et al. 2019

Ann Transl Med

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Background: Ulcerative colitis (UC) is an inflammatory bowel disease (IBD) that causes long-lasting inflammation and ulcers in the human digestive tract. The repair role of TLR4 in the intestinal epithelium is still unknown.Methods: By comparing to wild-type (WT) mice, Toll-like receptor 4 (TLR4)-knockout mice (TLR4-KO) were used as dextran sulfate sodium (DSS)-induced colitis models to explore the role of TLR4 signaling in intestinal injury. High-throughput RNA-Seq, RT-qPCR and ELISA were performed to screen and verify key differences in gut genes between WT and TLR4-KO mice. Functional study of core dysregulated factors was performed in intestinal cell lines.Results: We found that DSS-induced intestinal injury was aggravated by LPS (TLR4 agonist) and TLR4-KO. When compared to WT mice, IL6, CCL2, CSF3, IL11, Ccnb1, Ccnd1 and TNF-α significantly decreased and Fas and FasL have increased in the gut of TLR4-KO mice. IL6, CCL2, CSF3, Fas and FasL have all increased in CT-26 cells treated with LPS. Combined with the above data and KEGG enrichment, it can be assumed that TLR4-KO might aggravate DSS-induced intestinal damage by attenuating cell cycle, cytokinecytokine receptor interaction, and Toll-like receptor signaling pathway, and enhancing the apoptosis pathway.In the functional study of core dysregulated factors, it was found that LPS, IL6, IL11, CSF3, CCL2, S100A8, S100A9 and Mmp3 have improved viability of colon cancer cell lines and decreased apoptosis rate of mouse colon cancer cells when these were treated with DSS. However, Jo-2 (Fas agonistic monoclonal antibody) played the opposite role in colon cancer cells treated with DSS.Conclusions: TLR4 had a repairing effect on DSS-induced intestinal damage and it up-regulate IL6, CCL2 and CSF3. Fas and FasL enhanced DSS-induced colon injury in mice, but might have little to do with TLR4 signaling.

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Section: Quantitative Real-time Pcr (Qrt-pcr) and Enzymelinked Immunomentioning

confidence: 99%

Toll-like receptor 4 (TLR4) deficiency aggravates dextran sulfate sodium (DSS)-induced intestinal injury by down-regulating IL6, CCL2 and CSF3

Yunjie¹,

Hu²,

Zhao³

et al. 2019

Ann Transl Med

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“…RNA was reverse transcribed into cDNA using the BeyoRT™ First Strand cDNA Synthesis Kit (Beyotime Institute of Biotechnology). U6 was used as an endogenous control [ 16 ]. The primer sequences were as follows:…”

Section: Methodsmentioning

confidence: 99%

Echinacoside exerts anti-tumor activity via the miR-503-3p/TGF-β1/Smad aixs in liver cancer

Zhou

Zhang

et al. 2021

Cancer Cell Int

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Background Echinacoside (ECH) is the main active ingredient of Cistanches Herba, which is known to have therapeutic effects on metastatic tumors. However, the effects of ECH on liver cancer are still unclear. This study was to investigate the effects of ECH on the aggression of liver cancer cells. Methods Two types of liver cancer cells Huh7 and HepG2 were treated with different doses of ECH at different times and gradients. MTT and colony formation assays were used to determine the effects of ECH on the viability of Huh7 and HepG2 cells. Transwell assays and flow cytometry assays were used to detect the effects of ECH treatment on the invasion, migration, apoptosis and cell cycle of Huh7 and HepG2 cells. Western blot analysis was used to detect the effects of ECH on the expression levels of TGF-β1, smad3, smad7, apoptosis-related proteins (Caspase-3, Caspase-8), and Cyto C in liver cancer cells. The relationship between miR-503-3p and TGF-β1 was detected using bioinformatics analysis and Luciferase reporter assay. Results The results showed that ECH inhibited the proliferation, invasion and migration of Huh7 and HepG2 cells in a dose- and time-dependent manner. Moreover, we found that ECH caused Huh7 and HepG2 cell apoptosis by blocking cells in S phase. Furthermore, the expression of miR-503-3p was found to be reduced in liver tumor tissues, but ECH treatment increased the expression of miR-503-3p in Huh7 and HepG2 cells. In addition, we found that TGF-β1 was identified as a potential target of miR-503-3p. ECH promoted the activation of the TGF-β1/Smad signaling pathway and increased the expression levels of Bax/Bcl-2. Moreover, ECH could trigger the release of mitochondrial Cyto C, and cause the reaction Caspases grade. Conclusions This study demonstrates that ECH exerts anti-tumor activity via the miR-503-3p/TGF-β1/Smad aixs in liver cancer, and provides a safe and effective anti-tumor agent for liver cancer.

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“…Previous research has provided evidence to show a high expression of BTK in a variety of cancers [21][22][23][24]. BTK has also been considered a diagnostic biomarker and immunotherapeutic target for glioma patients [25,26].…”

Section: Introductionmentioning

confidence: 99%

MicroRNA-628-3p inhibits glioblastoma cell proliferation and GSC maintenance by directly targeting BTK

Wang

Cao

et al. 2022

Preprint

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Differentially expressed microRNAs (miRNAs) have been identified as a critical factor of glioblastoma (GBM). Therefore, we analyzed the pathogenic mechanism of a new miRNA (miR-628-3p) in glioblastoma. We identified miR-628-3p expression using qRT-PCR and FISH test, and a lot of in vitro tests looked at how miR-628-3p and BTK work biologically. Using a dual luciferase reporter experiment, the target site and mechanism of miR-628-3p binding to BTK were identified. Subcutaneous carcinogenesis was utilized to demonstrate the functionality of miR-628-3p in vivo. Inhibiting GBM growth and GSC maintenance was miR-628-3p overexpression. In addition, miR-628-3p regulated the expression of BTK, which was found as a promoter of GBM development in the research. Our findings showed that miR-628-3p reduces GBM growth and GSC maintenance, suggesting a potential therapeutic target for glioma treatment.

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Modeling qRT-PCR dynamics with application to cancer biomarker quantification

Cited by 8 publications

References 34 publications

Toll-like receptor 4 (TLR4) deficiency aggravates dextran sulfate sodium (DSS)-induced intestinal injury by down-regulating IL6, CCL2 and CSF3

Toll-like receptor 4 (TLR4) deficiency aggravates dextran sulfate sodium (DSS)-induced intestinal injury by down-regulating IL6, CCL2 and CSF3

Echinacoside exerts anti-tumor activity via the miR-503-3p/TGF-β1/Smad aixs in liver cancer

MicroRNA-628-3p inhibits glioblastoma cell proliferation and GSC maintenance by directly targeting BTK

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