Zearalenone exposure triggered porcine granulosa cells apoptosis via microRNAs-mediated focal adhesion pathway

The effects of ovine, porcine and human FSH, and ovine and human LH on the accumulation of cyclic AMP by porcine granulosa cells obtained from follicles at various stages of maturation were investigated. During incubation periods of 15 min, 10 μg ovine FSH pretreated with antiserum to LH or 10 μg human FSH resulted in an 11- to 18-fold, five-to ninefold, and less than a twofold increase in intracellular accumulation of cyclic AMP by granulosa cells from small (1–2 mm), medium (3–5 mm) and large (6–12 mm) follicles respectively. Similar patterns of response occurred with addition of porcine FSH. After incubation for 30 and 60 min with ovine, porcine or human FSH, significant accumulation of cyclic AMP in the incubation medium occurred with cells obtained from small and medium-sized follicles. After 60 min of incubation with FSH the accumulation of cyclic AMP in the incubation medium exceeded the intracellular cyclic AMP levels in granulosa cells from small and medium-sized follicles. During incubation periods of 15 min, 1·0 μg ovine LH resulted in less than a twofold, a fourfold and greater than a tenfold increase in intracellular accumulation of cyclic AMP by granulosa cells from small, medium and large follicles respectively. Addition of human LH brought about a similar response. Incubation periods of 30 and 60 min with 1·0 μg ovine or human LH resulted in significant accumulation of cyclic AMP in the incubation medium by granulosa cells from large follicles; cyclic AMP content in the incubation medium was greater after 60 min compared with 30 min of incubation. It was concluded that ovine FSH pretreated with an antiserum to LH had similar effects on cyclic AMP levels as did purified human and porcine FSH, and that the stimulatory effects of the less pure ovine FSH were probably not due to an impurity in the FSH preparation. Porcine granulosa cells obtained from small follicles should be suitable as an in-vitro FSH bioassay while granulosa cells obtained from large follicles should be suitable as an in-vitro LH bioassay.

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Regulation of Deoxyribonucleic Acid Synthesis in Cultured Porcine Granulosa Cells by Growth Factors and Hormones*

Endocrinology ◽

10.1210/endo-120-3-1039 ◽

1987 ◽

Vol 120 (3) ◽

pp. 1039-1046 ◽

Cited By ~ 37

Author(s):

JAMES M. HAMMOND ◽

HUGH F. ENGLISH

Keyword(s):

Growth Factors ◽

Granulosa Cells ◽

Deoxyribonucleic Acid ◽

Acid Synthesis ◽

Porcine Granulosa Cells ◽

Deoxyribonucleic Acid Synthesis

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Cytotoxic Effect of RecombinantMycobacterium tuberculosisCFP-10/ESAT-6 Protein on the Crucial Pathways of WI-38 Cells

Journal of Biomedicine and Biotechnology ◽

10.1155/2009/917084 ◽

2009 ◽

Vol 2009 ◽

pp. 1-10 ◽

Cited By ~ 7

Author(s):

Kun-Nan Tsai ◽

Err-Cheng Chan ◽

Tsung-Yeh Tsai ◽

Kuei-Tien Chen ◽

Chun-Yu Chen ◽

...

Keyword(s):

Focal Adhesion ◽

Microarray Data ◽

Cytotoxic Effect ◽

Time Course ◽

Essential Role ◽

Topological Properties ◽

Regulatory Effect ◽

Qrt Pcr ◽

Focal Adhesion Pathway ◽

Pathway Databases

To unravel the cytotoxic effect of the recombinant CFP-10/ESAT-6 protein (rCFES) on WI-38 cells, an integrative analysis approach, combining time-course microarray data and annotated pathway databases, was proposed with the emphasis on identifying the potentially crucial pathways. The potentially crucial pathways were selected based on a composite criterion characterizing the average significance and topological properties of important genes. The analysis results suggested that the regulatory effect of rCFES was at least involved in cell proliferation, cell motility, cell survival, and metabolisms of WI-38 cells. The survivability of WI-38 cells, in particular, was significantly decreased to 62% with 12.5 μMrCFES. Furthermore, the focal adhesion pathway was identified as the potentially most-crucial pathway and 58 of 65 important genes in this pathway were downregulated by rCFES treatment. Using qRT-PCR, we have confirmed the changes in the expression levels of LAMA4, PIK3R3, BIRC3, and NFKBIA, suggesting that these proteins may play an essential role in the cytotoxic process in the rCFES-treated WI-38 cells.

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