Authors:

Addison Hill

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About Paper:

Dairy cattle must synthesize 90% of their glucose requirements. Pyruvate carboxylase (PC) catalyzes the synthesis of oxaloacetate, which is a primary substrate for glucose production. Both saturated and unsaturated fatty acids regulate PC expression. The objective was to study the effect of altering fatty acids and PC expression in Madin-Darby Bovine Kidney (MDBK) cells on cell growth. We hypothesized that when PC expression is impaired by the addition of saturated fatty acids, or when PC is under-expressed, that cell growth would be impaired. MDBK cells were plated, grown overnight, and incubated for 21h in fatty acid treatments that totaled to 1mM, with varying ratios of palmitic acid to α-linolenic acid. Cells were counted to determine the total number and percent alive. In the second experiment, four cell lines were used: WT, PC shRNA scramble, PC shRNA knockdown, and PC overexpression. Cells were plated, grown overnight, and either treated with or without doxycycline. Cells were counted on d3, 5, and 7 for total cells and percent alive. Fatty acid treatment had an overall effect on total cells (P=0.001) and percent alive (P=0.0001), and the 1mM palmitic acid treatment reduced total cells and percent alive. There was a significant day effect on total cells and percent alive with d3 being different from d5 and 7 for total cells, but d5 not being different from d7. Overall, 1 mM palmitic acid appeared cytotoxic. PC knockdown did not impair cell growth, which suggests fatty acids control cell growth independent of PC.

Source:

Purdue University / 2023

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Co-authors:

Addison Hill