Regulation of miR-29b-1/a transcription and identification of target mRNAs in CHO-K1 cells

Muluhngwi, Penn; Richardson, Kirsten; Napier, Joshua; Rouchka, Eric C.; Mott, Justin L.; Klinge, Carolyn M.

doi:10.1016/j.mce.2017.01.044

Cited by 7 publications

(4 citation statements)

References 57 publications

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“…In our study, immunofluorescence analysis and transfection experiments using miRNA analogues suggested that these miRNAs might not only regulate genes that modulate the composition of ECM such as MMPs and TIMPs, but might also play a role in the alteration of hormonal activities via modifying the gene expression of estrogen and progesterone receptors. Consistent with our results, a few previous reports suggested that these miRNAs affect the receptivity of estrogen and progesterone receptors [ 43 , 44 , 45 , 46 , 47 ]. Our results suggest that miRNAs might play a role in the development of ULs via the regulation of the hormonal micro-environment and composition of ECM.…”

Section: Discussionsupporting

confidence: 93%

Variation in MicroRNA Expression Profile of Uterine Leiomyoma with Endometrial Cavity Distortion and Endometrial Cavity Non-Distortion

Kim

Shin

et al. 2018

IJMS

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The expression profile of microRNA (miRNA) in uterine leiomyoma (UL) cells is different from that in normal uterine myometrial (UM) cells. The effect of UL cells on uterine receptivity might vary according to their ability to distort the uterine endometrial cavity. However, the variation in miRNA expression profiles between endometrial cavity-distorting leiomyoma (ECDL) and endometrial cavity non-distorting leiomyoma (ECNDL) cells remains unknown. This study aimed to elucidate whether the expression profile of miRNAs in ECDL cells is dissimilar to that of ECNDL cells in uterus. Pelviscopic myomectomy was performed to obtain tissue samples of UL and their corresponding normal UM tissues (matched) from patients with UL (n = 26), among whom women with ECNDL and ECDL numbered 15 and 11, respectively. The relative expression of hsa-miR-15b, -29a, -29b, -29c, -197, and -200c as well as the candidate target genes in UL cells was compared to those in the matched UM cells using qRT-PCR to assess their ability to cause ECD. The spatial expression of miRNAs and target genes in the UL tissues was analyzed using in situ hybridization. Target gene expression was analyzed using qPCR after transfection with the mimics and inhibitors of miRNAs in UL cells. The relative expression level of miR-15b was upregulated, and the relative expression levels of miR-29a, -29b, -29c, -197, and -200c were downregulated in UL cells compared to those in UM cells. The relative expression levels of progesterone receptor, estrogen receptor, and matrix metalloproteinases (MMPs) were upregulated in UL cells compared to those in UM cells. The relative expression levels of miR-29c and -200c were downregulated, and the relative expression levels of estrogen receptor, MMPs and tissue inhibitors of metalloproteinases (TIMPs) were upregulated in ECDL cells compared to those in ECNDL cells. The expression profile of miRNAs in UL cells varied with respect to the occurrence or absence of endometrial cavity distortion. The biochemical properties of UL might be regulated by miRNAs in order to alter their effect on structural homeostasis of the uterus.

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Section: Discussionsupporting

confidence: 93%

Variation in MicroRNA Expression Profile of Uterine Leiomyoma with Endometrial Cavity Distortion and Endometrial Cavity Non-Distortion

Kim

Shin

et al. 2018

IJMS

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“…miR-29a is described in literature as upregulated in CHO cells during exponential growth phase (Klanert et al 2016 ), which is what we also observe as an enriched presence in the respective exosome samples. However, others also claim a growth-inhibitory effect of this miRNA for CHO cells (Bort et al 2012 ; Muluhngwi et al 2017 ). Similar applies to miR-378, which we found to be enriched in CHO exosomes and that is presumably growth-inhibitory species, since its depletion can increase growth and cell density of CHO cells (Coleman et al 2018 ; Costello et al 2018 ).…”

Section: Discussionmentioning

confidence: 99%

Exploring the molecular content of CHO exosomes during bioprocessing

Keysberg

Hertel

Schelletter

et al. 2021

Appl Microbiol Biotechnol

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In biopharmaceutical production, Chinese hamster ovary (CHO) cells derived from Cricetulus griseus remain the most commonly used host cell for recombinant protein production, especially antibodies. Over the last decade, in-depth multi-omics characterization of these CHO cells provided data for extensive cell line engineering and corresponding increases in productivity. However, exosomes, extracellular vesicles containing proteins and nucleic acids, are barely researched at all in CHO cells. Exosomes have been proven to be a ubiquitous mediator of intercellular communication and are proposed as new biopharmaceutical format for drug delivery, indicator reflecting host cell condition and anti-apoptotic factor in spent media. Here we provide a brief overview of different separation techniques and subsequently perform a proteome and regulatory, non-coding RNA analysis of exosomes, derived from lab-scale bioreactor cultivations of a CHO-K1 cell line, to lay out reference data for further research in the field. Applying bottom-up orbitrap shotgun proteomics and next-generation small RNA sequencing, we detected 1395 proteins, 144 micro RNA (miRNA), and 914 PIWI-interacting RNA (piRNA) species differentially across the phases of a batch cultivation process. The exosomal proteome and RNA data are compared with other extracellular fractions and cell lysate, yielding several significantly exosome-enriched species. Key points • First-time comprehensive protein and miRNA characterization of CHO exosomes. • Isolation protocol and time point of bioprocess strongly affect quality of extracellular vesicles. • CHO-derived exosomes also contain numerous piRNA species of yet unknown function.

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“…aberrant expression of miR-29b-1 and miR-29a reduced CHO-K1 cell proliferation, colony formation, and cell invasion. In addition, increased miR-29b-1/a expression reduced mRNA and protein levels in Disher ( Muluhngwi et al, 2017 ).…”

Section: Mechanism Of Mirna Affecting the Expression Of Recombinant P...mentioning

confidence: 99%

The Effect of microRNA on the Production of Recombinant Protein in CHO Cells and its Mechanism

Liu

Dong

Lin³

et al. 2022

Front. Bioeng. Biotechnol.

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Recombinant protein production by mammalian cells is the initial step in the manufacture of many therapeutic proteins. Chinese hamster ovary (CHO) cells are the most common host system to produce recombinant therapeutic proteins (RTPs). However, it is still challenging to maintain high productivity ensuring the good quality of RTPs produced by CHO cells. MicroRNAs(miRNAs) are short regulatory non-coding RNAs that can regulate cellular behavior and complex phenotypes. It has been found that miRNAs can enhance the expression level of recombinant proteins in CHO cells by promoting proliferation, resisting apoptosis, and regulating metabolism. miRNAs also can affect the quality of RTPs. In this review, we will discuss the effect and mechanism of miRNA on the expression level and quality of recombinant proteins in CHO cells.

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Regulation of miR-29b-1/a transcription and identification of target mRNAs in CHO-K1 cells

Cited by 7 publications

References 57 publications

Variation in MicroRNA Expression Profile of Uterine Leiomyoma with Endometrial Cavity Distortion and Endometrial Cavity Non-Distortion

Variation in MicroRNA Expression Profile of Uterine Leiomyoma with Endometrial Cavity Distortion and Endometrial Cavity Non-Distortion

Exploring the molecular content of CHO exosomes during bioprocessing

The Effect of microRNA on the Production of Recombinant Protein in CHO Cells and its Mechanism

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