The serum- and glucocorticoid-inducible kinase-1 (SGK1) is ubiquitously expressed and under genomic control by cell stress (including cell shrinkage) and hormones (including gluco- and mineralocort...

https://www.physiology.org/doi/pdf/10.1152/physrev.00050.2005

(Patho)physiological Significance of the Serum- and Glucocorticoid-Inducible Kinase Isoforms

mTOR-Raptor Binds and Activates SGK1 to Regulate p27 Phosphorylation

Dihydrotestosterone-inducible dickkopf 1 from balding dermal papilla cells causes apoptosis in follicular keratinocytes.

Serum and glucocorticoid inducible protein kinase (SGK) plays a crucial role in promoting cell survival, but the mechanisms for this response are not clear. We show that SGK is involved in the regulation of apoptosis in breast cancer cells by modulating the transcriptional activity of nuclear transcription factor kappaB (NF-kappaB). High levels of SGK expression were observed in human breast cancer samples. When SGK was reduced the apoptotic rate increased, and increased SGK activity prevents serum withdrawal-induced apoptosis. SGK-induced cell survival was abolished by a dominant-negative form of IkappaB kinase beta (IKKbeta, K44A) or a null mutation of IKKbeta in mouse embryonic fibroblast cells indicating involvement of the NF-kappaB pathway. Serum-induced SGK or increased expression of SGK activated NF-kappaB transcriptional activity, whereas small interference RNA to SGK blocked NF-kappaB activity. Coexpression of SGK and IKKbeta significantly increased the activation of NF-kappaB (versus expression of IKKbeta alone). Expression of dominant-negative IKKbeta K44A, IkappaBalpha AA, and kinase-dead SGK (127KM) blocked the ability of SGK to stimulate NF-kappaB activity, suggesting that IKKbeta is a target of SGK. We also show that SGK enhances the ability of IKKbeta to phosphorylate endogenous IkappaBalpha in cells or recombinant glutathione S-transferase-IkappaBalpha in vitro and increases IkappaBalpha degradation; SGK physically associates with and activates IKKbeta in MDA231 cells via phosphorylation of Ser(181) in IKKbeta. Taken together, we conclude that SGK acts as an oncogene in breast cancer cells through activation of the IKK-NF-kappaB pathway, thereby preventing apoptosis. Blocking SGK expression/activity represents a potential therapeutic approach for breast cancer treatment.

https://cancerres.aacrjournals.org/content/canres/65/2/457.full.pdf

Antiapoptotic Effect of Serum and Glucocorticoid-Inducible Protein Kinase Is Mediated by Novel Mechanism Activating IκB Kinase

The completely assembled human genome has made it possible for modern medicine to step into an era rich in genetic information and high-throughput genomic analysis. These novel and readily available genetic resources and analytical tools may be the key to unravel the molecular basis of hepatocellular carcinoma (HCC). Moreover, since an efficient treatment for this disease is lacking, further understanding of the genetic background of HCC will be crucial in order to develop new therapies aimed at selected targets. We report on the current status and recent developments in HCC genetics. Special emphasis is given to the genetics and regulation of major signalling pathways involved in HCC such as p53, Wnt-signalling, TGFβ, Ras, and Rb pathways. Furthermore, we describe the influence of chromosomal aberrations as well as of DNA methylation. Finally, we report on the rapidly developing field of genomic expression profiling in HCC, mainly by microarray analysis.

Genetics of hepatocellular carcinoma.

We performed gene expression profiling of normal and hepatocellular carcinoma (HCC) liver tissues using a high-density microarray that contained 3,063 human cDNA. The results of a microarray hybridization experiment from eight different HCC tissues were analyzed and classified by the Cluster program. Among these differentially-expressed genes, the galectin-3, serine/threonine kinase SGK, translation factor eIF-4A, -4B, -3, fibroblast growth factor receptor, and ribosomal protein L35A were up-regulated; the mRNAs of Nip3, decorin, and the insulin-like growth factor binding protein-3 were down-regulated in HCC. The differential expression of these genes was further confirmed by an RT-PCR analysis. In addition, our data suggest that the gene expression profile of HCC varies according to the histological types.

Gene expression profile analysis in human hepatocellular carcinoma by cDNA microarray.

Nopaline synthase (nos) is of bacterial origin and expressed in plant tissues to generate an unusual compound, nopaline, which is secreted into the environment where Agrobacterium tumefaciens uses it as a nutrient source. The nos promoter contains three distinctive regions which are important for the promoter activity. Among these, the upstream region between -131 and -112 is essential for the nos promoter. The upstream region consists of two hexamer motifs which are separated by an eight nucleotide sequence. In this study, we have investigated the role of the hexamer motif that is located between -117 and -112. Increasing the distance between the hexamer and the downstream regulatory region significantly reduced the promoter activity, indicating that a proper distance between the regulatory elements is needed for an optimum level of activity. Insertion of a single copy of the hexamer enhanced the promoter in both orientations. Point mutations in the inserted hexamer sequence reduced the enhancing effect. These results confirmed that the hexamer motif is important for the nos promoter activity. The b sequence that is located immediately downstream of the hexamer motif did not function by itself, but together with the hexamer it appeared to enhance the promoter activity. These indicate that the sequence surrounding of the upstream regulatory element is also involved in controlling the promoter function.

Characterization of a hexamer motif and b element of the nopaline synthase (nos) promoter.

The flavonoid 3′,5′-hydroxylase (F3′,5′H) gene is in the NADPH-Cytochrome P450 reductase family. A partial F3′,5′H cDNA was cloned by RT-PCR from corollas ofGentiana scabra. Activity of F3′,5′H was observed during flower development, and activity was highest at the stage prior to flower opening. The enzyme activity was increased with the addition of SO μM gibberellic acid (GA3) immediately prior to flower opening. Western blot analysis showed that purified F3′,5′H protein fromG. scabra was cross-reacted with rabbit anti-Gentiana triflora NADPH-Cyt P450 reductase antibodies. Only one 80 to 82 kDa protein was detected by western blot analysis of fresh microsomal extracts. The enzyme activity was not distinctly changed by adding 50 μM GA3 to corollas. Therefore, we conclude that GA3 controls the expression of the F3′,5′H gene at the transcriptional level, not at the translational level.

Gibberellic acid regulates flavonoid 3′,5′-hydroxylase gene transcription in the corolla ofGentiana scabra

Lipases are useful enzymes that are primarily responsible for the hydrolysis of acylglycerides during lipid processing. We have cloned a lipase gene from a rice seed coat cDNA library (Oryza sativa cv. Dongjin). The cDNA was 1,445 bp in length and encoded 361 amino acid residues (GenBank accession No. AY580163). The deduced amino acid sequence had 82 and 56% identity to Oryza sativa (cv. Chuchung) and Arabidopsis thaliana lipase genes, respectively, and there was a GxSxG consensus motif near the catalytic triad at the active serine site. The deduced sequence had little homology to mammalian and microbial lipases. When the Oryza sativa lipase gene was expressed in Escherichia coli with the pET expression system, activity was found mainly in the pellet fraction. The purified product had lipolytic activity towards tributyrin and was about 40 kDa in size.

Cloning and Expression of a Lipase Gene from Rice (Oryza sativa cv. Dongjin)

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Papers

Gene expression profile analysis in human hepatocellular carcinoma by cDNA microarray.

Characterization of a hexamer motif and b element of the nopaline synthase (nos) promoter.

Gibberellic acid regulates flavonoid 3′,5′-hydroxylase gene transcription in the corolla ofGentiana scabra

Cloning and Expression of a Lipase Gene from Rice (Oryza sativa cv. Dongjin)