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Study of transcriptional changes in MCM7-depleted and G1/S-arrested A549 and A375 cells

ABSTRACT: We tested if the changes in cell invasion and migration observed after MCM7 depletion manifested in an altered gene expression profile. For this purpose, we performed RNAseq analysis after MCM7 knock-down in A549 and A375 cells expressing a Dox-inducible shMCM7. As controls, we used shMCM7 cells that had not been induced with Dox, as well as the parental A549 and A375 cell lines grown in the presence or absence of Dox. To exclude possible effects caused by changes in cell cycle progression, all RNA samples were extracted from double thymidine synchronized cell populations, in which >98% cells were arrested at the G1/S phase boundary.

ORGANISM(S): Homo sapiens

PROVIDER: GSE149523 | GEO | 2021/01/01

REPOSITORIES: GEO

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Project description:miR-93/106b and their host gene minichromosome maintenance complex component 7 (MCM7) reside at chr7q22, a region frequently rearranged in leiomyomas. We explored the expression of miR-93/106b in leiomyoma and paired myometrium (N=62) from untreated and patients exposed to hormonal therapies (GnRHa, Depo-Provera and oral contraceptives) from African Americans and Caucasians, and their regulatory functions in isolated paired (N=15) leiomyoma and myometrial smooth muscle cells (LSMC and MSMC) and leiomyosarcoma cell line (SKLM-S1). At tissue level leiomyomas expressed significantly lower levels of miR-93 and elevated MCM7 as compared to myometrium with limited racial influence or hormonal exposure on their expression. Assessing the regulatory function of miR-93/106b through doxycycline-inducible lentiviral transduction in microarray analysis, tissue factor (F3) and IL-8 were identified as their possible targets. At tissue level leiomyomas expressed a significantly lower level of F3 and an elevated IL-8 which exhibited an inverse relationship with miR-93, but with limited racial or hormonal influences. Gain-of-function of miR-93/106b in LSMC, MSMC and SKLM-S1 dose-dependently repressed F3 and IL-8 through direct interactions with their respective 3M-bM-^@M-^YUTRs and indirectly through F3 repression inhibited IL8, CTGF and PAI-1 expression, confirmed by using siRNA silencing or factor Vlla (FVIIa) activation of F3, as well as reducing the rate of proliferation, while increasing caspase 3/7 activity. We concluded that differential expression of miR-93/106b and their direct and/or indirect regulatory functions on F3, IL-8, CTGF and PAI-1 expression, with key roles in inflammation and tissue turnover may be of significance in the outcome of leiomyoma growth and associated symptoms. Total RNA isolated from TF324 cells transfected with DOX-inducible lentiviral construct carrying miR-106b~25 cluster with and without Dox treatments for 6 days was subjected to gene expression profiling using Sentirx Beadchip Array HumanHT-12_v4.

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Study of transcriptional changes in MCM7-depleted and G1/S-arrested A549 and A375 cells

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