casein-kinase-ii and Epilepsy--Temporal-Lobe

To investigate mechanisms that TMEM2 activation inhibits hepatitis B virus (HBV) infection in hepatocarcinoma (HCC) cells, co-immunoprecipitation (Co-IP) and mass spectrometry were used in screening interacting proteins for TMEM2. Levels of casein kinase 2 subunit α3 (CSNK2A3) in HCC cells were found to be inhibited or overexpressed using siRNAs and pcDNA3.1-CSNK2A3, respectively. Effect of CSNK2A3 expression on cell proliferation was analyzed using MTS, while its effect on HBV infection was measured using ddPCR and IHC. Western blotting and JAK inhibitor ruxolitinib were also used to determine whether TMEM2-regulated CSNK2A3 expression and HBV infection were affected by JAK-STAT signaling. Co-IP and mass spectrometry results showed that CSNK2A3 interacts with TMEM2. Moreover, overexpression of CSNK2A3 significantly inhibited cell proliferation, while inhibition of CSNK2A3 promoted proliferation of HCC cells. In addition, overexpression of CSNK2A3 was observed to significantly enhance HBV infection, while siRNA knockdown of CSNK2A3 inhibited HBV infection. Notably, effect of CSNK2A3 overexpression on HBV infection was suppressed by TMEM2 overexpression. Further mechanistic analyses have revealed that TMEM2 could antagonize the effects of CSNK2A3 on cell proliferation and HBV infection via JAK-STAT pathway activation. In conclusion, TMEM2 has been determined to bind to CSNK2A3 to inhibit HBV infection via activation of the JAK-STAT pathway.

Topics: Brain; Carcinoma, Hepatocellular; Case-Control Studies; Casein Kinase II; Epilepsy, Temporal Lobe; Hepatitis B; Hepatitis B virus; Humans; Janus Kinase 1; Liver Neoplasms; Membrane Proteins; PPAR gamma; STAT Transcription Factors; Tumor Cells, Cultured

The slow afterhyperpolarizing potential (sAHP) following prolonged depolarization is a major intrinsic mechanism of neuronal inhibition, by powerfully dampening excitability for up to 2 s. Therefore, an altered sAHP function might be vulnerable to hyperexcitable states such as epilepsy. Here, we have investigated the role of casein kinase 2 (CK2) on the sAHP in control and chronically epileptic tissue.. Using the rat pilocarpine model of chronic temporal lobe epilepsy, we performed whole-cell patch-clamp recordings of acutely isolated CA1 pyramidal cells and field potential measurements on hippocampal slices.. Chronic oral administration of the CK2 inhibitor 4,5,6,7-tetrabromotriazole (TBB) for 4 days prior to brain dissection caused a significant increase of the sAHP-mediating current in both control and epileptic tissues. In contrast, when TBB was acutely applied during the patch-clamp recording, the sAHP remained unaltered, indicating that chronic CK2 inhibition was required for sAHP augmentation. To test whether CK2 inhibition also has an anticonvulsive effect, we evoked recurrent epileptiform discharges (REDs) in hippocampal slice preparations by Mg²⁺ removal. It is important to note that chronic oral TBB administration abolished REDs induced by 0-Mg²⁺ solution, suggesting that CK2 inhibition indeed has anticonvulsive and perhaps antiepileptogenic properties.. Our data demonstrated that CK2 inhibition augments the sAHP and might represent a novel mechanism of action of anticonvulsant drugs.

Topics: Animals; Anticonvulsants; CA1 Region, Hippocampal; Casein Kinase II; Disease Models, Animal; Epilepsy, Temporal Lobe; Hydrocarbons, Brominated; Male; Membrane Potentials; Patch-Clamp Techniques; Pilocarpine; Rats; Rats, Wistar; Triazoles