Project description:An increasing number of studies have shown that long noncoding RNA (lncRNA) dysregulation plays an important role in development of various cancers, including colon cancer. Nonetheless, the potential mechanisms of lncRNA in regorafenib-resistance remain unclear. Our research revealed the lncRNA AC069513.3 (MIR570MG) increased in regorafenib-resistant colon cancer cells compared to the regorafenib-sensitive cells. Furthermore, lncRNA AC069513.3 (MIR570MG) sponged miR-145, which declined in regorafenib-resistant colon cancer cell lines. More importantly, overexpression of miR-145 hampered cell proliferation and retrieved colon cancer regorafenib-sensitivity, contrary to the function of lncRNA AC069513.3 (MIR570MG). Dual luciferase reporter assay confirmed that miR-145 bound to 3’-UTR of SMAD3, a transcriptional modulator activated by TGFβ, resulting in blockage of TGFβ /SMAD3-mediated cell growth and cycle progression. Furthermore, ectopic expression of miR-145 inhibitor in the parental cells endowed resistance to regorafenib. Conversely, knockdown of AC069513.3 (MIR570MG) impoverished resistance against regorafenib. In summary, our findings suggested that lncRNA AC069513.3 (MIR570MG) promoted regorafenib resistance via releasing SMAD3 from miR-145, leading to activation of SMAD3-mediated signaling pathways. Long noncoding RNA profiling by RT-PCR
Project description:mRNA levels of the known 91 FoxO1 target genes were evaluated with RT2 Profiler PCR array in cardiomyocytes transduced with LacZ, Mst1, FoxO1, FoxO1+Mst1, or FoxO1+DN-Mst1 genes were evaluated with RT2 Profiler PCR arrays.
Project description:Wild type (WT), Nod2-/-, Pglyrp3-/-, and Pglyrp3-/-Nod2-/- mice (BALB/c) were treated with oral 5% dextran sodium sulfate (DSS) for 48, 72, or 96 hrs, their colons were removed and homogenized, and RNA was isolated using the TRIZOL method. Quantitative reverse transcription real-time PCR (qRT-PCR) was used to quantify the amounts of mRNA in the colon using the inflammatory gene expression RT2 Profiler PCR Array from Qiagen/SA Biosciences.
Project description:LncRNAs have been recently implicated as having oncogenic and tumor suppressor roles.To further investigate the function of lncRNA in colon cancer, we have employed lncRNA microarray to identify lncRNA with the potential to involve the metastasis of colon cancer. Human peripheral blood from healthy donors was irradiated ex vivo, and a 74-gene consensus signature was identified that distinguished between four radiation doses (0.5, 2, 5 and 8 Gy) and control samples. The same set of genes separated samples by exposure level at both six and 24 hours after treatment, with overlap evident only at the highest two doses (5 and 8 Gy). Expression of five genes (CDKN1A, FDXR, SESN1, BBC3 and PHPT1) from this signature was quantified in the same RNA samples by real-time PCR, confirming low variability between donors as well as the predicted radiation response pattern. The experimental samples are divided into three groups(normalM-cM-^@M-^Aprimary tumor and metastatic tumor) to compare lncRNA expression profiling of those
Project description:Wild type (WT), Nod2-/-, Pglyrp3-/-, and Pglyrp3-/-Nod2-/- mice (BALB/c) were treated with oral 5% dextran sodium sulfate (DSS) for 48, 72, or 96 hrs, their colons were removed and homogenized, and RNA was isolated using the TRIZOL method. Quantitative reverse transcription real-time PCR (qRT-PCR) was used to quantify the amounts of mRNA in the colon using the inflammatory gene expression RT2 Profiler PCR Array from Qiagen/SA Biosciences. qRT-PCR gene expression profiling
Project description:LncRNAs have been recently implicated as having oncogenic and tumor suppressor roles.To further investigate the function of lncRNA in colon cancer, we have employed lncRNA microarray to identify lncRNA with the potential to involve the metastasis of colon cancer. Human peripheral blood from healthy donors was irradiated ex vivo, and a 74-gene consensus signature was identified that distinguished between four radiation doses (0.5, 2, 5 and 8 Gy) and control samples. The same set of genes separated samples by exposure level at both six and 24 hours after treatment, with overlap evident only at the highest two doses (5 and 8 Gy). Expression of five genes (CDKN1A, FDXR, SESN1, BBC3 and PHPT1) from this signature was quantified in the same RNA samples by real-time PCR, confirming low variability between donors as well as the predicted radiation response pattern.