Reprogramming progressive cells display low CAG promoter activity

AbstractThere is wide variability in the propensity of somatic cells to reprogram into pluripotency in response to the Yamanaka factors. How to segregate these variability to enrich for cells of specific traits that reprogram efficiently remains challenging. Here we report that the variability in reprogramming propensity is associated with the activity of the MKL1/SRF transcription factor and concurs with small cell size as well as rapid cell cycle. Reprogramming progressive cells can be prospectively identified by their low activity of a widely used synthetic promoter, CAG. CAGlow cells arise and expand during cell cycle acceleration in the early reprogramming culture of both mouse and human fibroblasts. Our work illustrate a molecular scenario underlying the distinct reprogramming propensities and demonstrate a convenient practical approach for their enrichment.

Download Full-text

ID1 mediates resistance to osimertinib in EGFR T790M-positive non-small cell lung cancer through epithelial–mesenchymal transition

BMC Pulmonary Medicine ◽

10.1186/s12890-021-01540-4 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Kejun Liu ◽

Xianwen Chen ◽

Ligang Wu ◽

Shiyuan Chen ◽

Nianxin Fang ◽

...

Keyword(s):

Lung Cancer ◽

Cell Cycle ◽

Small Cell Lung Cancer ◽

Cell Line ◽

Epithelial Mesenchymal Transition ◽

Small Cell ◽

Small Cell Lung ◽

Mesenchymal Transition ◽

Egfr T790m ◽

E Cadherin

Abstract Background ID1 is associated with resistance to the first generation of EGFR tyrosine kinase inhibitors (EGFR-TKIs) in non-small cell lung cancer (NSCLC). However, the effect of ID1 expression on osimertinib resistance in EGFR T790M-positive NSCLC is not clear. Methods We established a drug-resistant cell line, H1975/OR, from the osimertinib-sensitive cell line H1975. Alterations in ID1 protein expression and Epithelial–mesenchymal transition (EMT)-related proteins were detected with western blot analysis. RT-PCR was used to evaluate the differences of gene mRNA levels. ID1 silencing and overexpression were used to investigate the effects of related gene on osimertinib resistance. Cell Counting Kit-8 (CCK8) was used to assess the proliferation rate in cells with altered of ID1 expression. Transwell assay was used to evaluate the invasion ability of different cells. The effects on the cell cycle and apoptosis were also compared using flow cytometry. Results In our study, we found that in osimertinib-resistant NSCLC cells, the expression level of the EMT-related protein E-cadherin was lower than that of sensitive cells, while the expression level of ID1 and vimentin were higher than those of sensitive cells. ID1 expression levels was closely related to E-cadherin and vimentin in both osimertinib-sensitive and resistant cells. Alteration of ID1 expression in H1975/OR cells could change the expression of E-cadherin. Downregulating ID1 expression in H1975/OR cells could inhibit cell proliferation, reduce cell invasion, promote cell apoptosis and arrested the cell cycle in the G1/G0 stage phase. Our study suggests that ID1 may induce EMT in EGFR T790M-positive NSCLC, which mediates drug resistance of osimertinib. Conclusions Our study revealed the mechanism of ID1 mediated resistance to osimertinib in EGFR T790M-positive NSCLC through EMT, which may provide new ideas and methods for the treatment of EGFR mutated NSCLC after osimertinib resistance.

Download Full-text

Alu RNA Modulates the Expression of Cell Cycle Genes in Human Fibroblasts

International Journal of Molecular Sciences ◽

10.3390/ijms20133315 ◽

2019 ◽

Vol 20 (13) ◽

pp. 3315 ◽

Cited By ~ 3

Author(s):

Simona Cantarella ◽

Davide Carnevali ◽

Marco Morselli ◽

Anastasia Conti ◽

Matteo Pellegrini ◽

...

Keyword(s):

Cell Cycle ◽

Cell Cycle Progression ◽

Human Fibroblasts ◽

Rna Polymerase Iii ◽

Protein Coding ◽

Non Coding Rna ◽

Genome Wide ◽

Hela Cell Lines ◽

Significant Enrichment ◽

Alu Rna

Alu retroelements, whose retrotransposition requires prior transcription by RNA polymerase III to generate Alu RNAs, represent the most numerous non-coding RNA (ncRNA) gene family in the human genome. Alu transcription is generally kept to extremely low levels by tight epigenetic silencing, but it has been reported to increase under different types of cell perturbation, such as viral infection and cancer. Alu RNAs, being able to act as gene expression modulators, may be directly involved in the mechanisms determining cellular behavior in such perturbed states. To directly address the regulatory potential of Alu RNAs, we generated IMR90 fibroblasts and HeLa cell lines stably overexpressing two slightly different Alu RNAs, and analyzed genome-wide the expression changes of protein-coding genes through RNA-sequencing. Among the genes that were upregulated or downregulated in response to Alu overexpression in IMR90, but not in HeLa cells, we found a highly significant enrichment of pathways involved in cell cycle progression and mitotic entry. Accordingly, Alu overexpression was found to promote transition from G1 to S phase, as revealed by flow cytometry. Therefore, increased Alu RNA may contribute to sustained cell proliferation, which is an important factor of cancer development and progression.

Download Full-text

Biocompatible micro-sized cell culture chamber for the detection of nanoparticle-induced IL8 promoter activity on a small cell population

Nanoscale Research Letters ◽

10.1186/1556-276x-6-505 ◽

2011 ◽

Vol 6 (1) ◽

pp. 505 ◽

Cited By ~ 10

Author(s):

Yvonne Kohl ◽

Gertie J Oostingh ◽

Adam Sossalla ◽

Albert Duschl ◽

Hagen von Briesen ◽

...

Keyword(s):

Cell Culture ◽

Cell Population ◽

Promoter Activity ◽

Small Cell ◽

Culture Chamber

Download Full-text

Amentoflavone Induces Cell-cycle Arrest, Apoptosis, and Invasion Inhibition in Non-small Cell Lung Cancer Cells

Anticancer Research ◽

10.21873/anticanres.14893 ◽

2021 ◽

Vol 41 (3) ◽

pp. 1357-1364

Author(s):

WEI-TING CHEN ◽

CHENG-HSIEN CHEN ◽

HUNG-TAI SU ◽

PO-FU YUEH ◽

FEI-TING HSU ◽

...

Keyword(s):

Lung Cancer ◽

Cell Cycle ◽

Small Cell Lung Cancer ◽

Cell Cycle Arrest ◽

Cancer Cells ◽

Cell Lung Cancer ◽

Small Cell ◽

Small Cell Lung ◽

Cycle Arrest ◽

Invasion Inhibition

Download Full-text

Alantolactone pyrazoline analogue inhibits cancer cell proliferation and induces apoptosis in non-small cell lung carcinoma cells

Bangladesh Journal of Pharmacology ◽

10.3329/bjp.v10i2.22619 ◽

2015 ◽

Vol 10 (2) ◽

pp. 409 ◽

Cited By ~ 4

Author(s):

Jing Lv ◽

Ming-Qin Cao ◽

Jian-Chun Yu

Keyword(s):

Cell Cycle ◽

Flow Cytometry ◽

Cell Cycle Arrest ◽

Chromatin Condensation ◽

Small Cell ◽

Small Cell Lung ◽

Cell Lung Carcinoma ◽

Cycle Arrest ◽

H460 Cells ◽

Dose Dependent

<p>The aim of the current study was to evaluate the anticancer and apoptotic effects of alantolactone pyrazoline analogue in human non-small cell lung cancer (NCI-H460) cells. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetra-zolium bromide) assay was used to evaluate the cell viability while as fluorescence microscopy was used to assess the effect on apoptosis, cellular and nuclear morphology. Flow cytometry evaluated the effect of APA on cell cycle arrest in these cells. The results revealed that APA induced potent, time and dose-dependent cytotoxic effects on the growth of NCI-H460 cells. It also inhibited colony forming tendency as well as cell invasion capability of these cancer cells. APA induced dose-dependent nuclear and cellular morphological effects including chromatin condensation and DNA fragmentation. Flow cytometry revealed that the anticancer effects of APA might be due to its cell cycle arrest inducing tendency in G0/G1 phase of the cell cycle.</p>

Download Full-text

Gastric Adenocarcinoma Predictive Long Intergenic Non-Coding RNA Promotes Tumor Occurrence and Progression in Non-Small Cell Lung Cancer via Regulation of the miR-661/eEF2K Signaling Pathway

Cellular Physiology and Biochemistry ◽

10.1159/000495831 ◽

2018 ◽

Vol 51 (5) ◽

pp. 2136-2147 ◽

Cited By ~ 6

Author(s):

Haiting Gu ◽

Junfeng Chen ◽

Yukang Song ◽

Haiyan Shao

Keyword(s):

Lung Cancer ◽

Cell Cycle ◽

Cell Proliferation ◽

Western Blot ◽

Small Cell Lung Cancer ◽

Small Cell ◽

Luciferase Reporter ◽

Small Cell Lung ◽

Non Coding Rna

Background/Aims: Long non-coding RNAs (lncRNAs) play vital roles in carcinogenesis as oncogenes or tumor suppressor genes. This study explored the biological function of lncRNA gastric adenocarcinoma predictive long intergenic non-coding RNA (GAPLINC) in human non-small cell lung cancer (NSCLC). Methods: GAPLINC expression in NSCLC specimens and cell lines was detected by qRT-PCR and Western blot. The effect of GAPLINC on cell proliferation was investigated using CCK8-assay, colony formation assay, and xenograft model. The effects of GAPLINC on apoptosis and cell cycle were determined using flow cytometry. The mechanism of GAPLINC involved in NSCLC was explored using Western blot, luciferase reporter assay, and RNA fluorescence in situ hybridization. Results: We found that GAPLINC expression was up-regulated in NSCLC tissues and cell lines. Overexpression of GAPLINC was associated with poor prognosis in patients with NSCLC. Silencing of GAPLINC significantly inhibited cell proliferation, promoted apoptosis, and induced cell cycle arrest in the G0/G1 phase. Results from xenograft transplantation showed that GAPLINC silencing inhibited the tumor growth in vivo. Interestingly, GAPLINC silencing decreased the expression of eukaryotic elongation factor-2 kinase (eEF2K) protein both in vivo and in vitro. Bioinformatic analysis and luciferase reporter confirmed that miR-661 targeted GAPLINC and eEF2K 3’-UTR and was negatively correlated with the expression of GAPLINC and eEF2K. Conclusion: Our findings indicate that GAPLINC promotes NSCLC tumorigenesis by regulating miR-661/eEF2K cascade and provide new insights for the pathogenesis underlying NSCLC and potential targets for therapeutic strategy.

Download Full-text