Articles
The effect of PLA2G6 differential expression on apoptosis and endoplasmic reticulum stress-related factors in hippocampal neurons
Abstract
Objective To explore the effects of PLA2G6 expression differences on hippocampal neuron apoptosis and endoplasmic reticulum stress-related factors. Methods The mouse hippocampal neuronal cell line HT22 was divided into five groups: blank control group (WT group), PLA2G6 overexpression group (OE group), overexpression control group (OE-ctrl group), PLA2G6 knockout group (KO group), and knockout control group (KO-ctrl group). The WT group received no treatment. The OE and OE-ctrl groups were transfected with PLA2G6 overexpression lentivirus and control lentivirus respectively, followed by puromycin screening to establish stable cell lines. RT-qPCR was used to detect the mRNA expression of PLA2G6 in each group. Western blot was applied to determine the protein expression levels of iPLA2, Bax, Bcl-2, PERK, IRE1 and ATF4 in HT22 cells. Results ① Fluorescence microscopy observation showed that cells in the OE and OE-ctrl groups were in good condition with high fluorescence abundance. ② Compared with the WT and OE-ctrl groups, the mRNA expression level of PLA2G6 was significantly increased in the OE group (P<0.05); compared with the WT and KO-ctrl groups, the mRNA expression level of PLA2G6 was markedly decreased in the KO group (P<0.05). ③ The protein expression of iPLA2 was significantly upregulated in the OE group versus the WT and OE-ctrl groups (P<0.05), while obviously downregulated in the KO group versus the WT and KO-ctrl groups (P<0.05). ④ In the OE group, Bax protein expression showed no significant difference compared with the WT and OE-ctrl groups (P>0.05); Bcl-2 protein expression was significantly elevated (P<0.05), and the Bax/Bcl-2 ratio was notably reduced (P<0.05). ⑤ The protein levels of PERK, IRE1 and ATF4 were significantly decreased in the OE group relative to the WT and OE-ctrl groups (P<0.05). Conclusion Stable HT22 cell lines with PLA2G6 overexpression and knockout are successfully constructed. Overexpression of PLA2G6 can downregulate the expression of apoptosis and endoplasmic reticulum stress-related indicators, whereas PLA2G6 knockout exerts no significant effect on these indicators.
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References
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