Localization of Prohibitin in the Nuclear Matrix and Alteration of Its Expression During Differentiation of Human Neuroblastoma SK-N-SH Cells Induced by Retinoic Acid

Abstract

The nuclear matrix-intermediate filament system of human neuroblastoma SK-N-SH cells before and after retinoic acid (RA) treatment was selectively extracted and the distribution of prohibitin (PHB) in the nuclear matrix, as well as its colocalization with related genes, was observed. Results of two-dimensional gel electrophoresis (2-DE), mass spectrometry (MS) identification, and protein immunoblotting all confirm that PHB was present in the components of SK-N-SH nuclear matrix proteins and was down-regulated after RA treatment. Immunofluorescence microscopy observations show that PHB was localized in the nuclear matrix and its distribution was altered due to RA treatment. Laser confocal microscopy results reveal that PHB colocalized with the expression products of c-myc, c-fos, p53, and Rb, but the colocalization region was altered after RA treatment. Our results prove that PHB is a nuclear matrix protein and is localized in nuclear matrix fibers. The distribution of PHB in SK-N-SH cells and its colocalization with related proto-oncogenes and tumor suppressor genes suggest that PHB plays pivotal roles in the differentiation of SK-N-SH cells and deserves further study.

All-Trans Retinoic Acid Treatment Is Associated with Prohibitin Expression in Renal Interstitial Fibrosis Rats

“…Li et al . [9] showed that prohibitin was localized in the nuclear matrix in human neuroblastoma SK-N-SH cells and its distribution was altered due to retinoic acid treatment, but they did not explore whether the retinoic acid treatment was associated with prohibitin expression.…”

“…It plays a pivotal role in the processes of cell differentiation and apoptosis [9]. The over-expression of prohibitin could protect the mitochondria from ROS-induced injury [10].…”

β-catenin induces expression of prohibitin gene in acute leukemic cells

“…PHB was distributed mainly in the regions of the nuclear membrane and cytoplasm in several cell types (45). Also PHB exists as a component of the nuclear matrix, and expression of PHB was downregulated during differentiation (27,46). In SK-N-SH cells, the expression of PHB is relatively strong compared with retinoic acid-treated cells.…”

“…In a B-cell lymphoma line, overexpression of PHB protects the cells from apoptosis, via regulation of E2F activity (26). Also retinoic acid or hexamethylene bisacetamide can stimulate the translocation of PHB to the nucleus in tumor cells (27). …”

Relationship Between the Prohibitin 3' Untranslated Region C > T Gene Polymorphism and Cancer Susceptibility - Results of a Meta-analysis

“…Early diagnosis and better prognosis of breast and ovarian cancers is still a challenge, and genetic factors have established a role in pathogenesis of breast and ovarian cancers (Yip et al, 2009;Gao et al, 2012;Jeschke et al, 2012;. Prohibitin can co-localize with the expressions of tumor suppressor genes, such as c-myc, c-fos, p53, and Rb (Li et al, 2011). Interestingly, c-myc, c-fos, p53, and Rb are very important in the pathogenesis of breast and ovarian cancers (Wang et al, 2010;Alshatwi et al, 2011;Kermani et al, 2011;Luparello et al, 2012;Szabova et al, 2012;Vijayaraman et al, 2012).…”