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Objective:To investigate the relationship between cervical lesion development and histone acetylation that regulates RAR-β2 expression. Methods:Immunohistochemistry and Western blot analysis were performed to detect AcH3, RAR-β2, and involu-crin expression in normal cervical tissues as well as in tissues with cervical intraepithelial neoplasia (CIN)Ⅱ-Ⅲand squamous cell cer-vical carcinoma. The relationship among histone acetylation level, RAR-β2 expression, and cervical lesion severity were analyzed. Re-sults:AcH3, RAR-β2, and involucrin expression were reduced or absent with the progression of cervical lesions;significant differences were noted between the groups (P<0.05). Histone acetylation level and RAR-β2 expression were positively correlated (r=0.797, P<0.05). AcH3 and RAR-β2 expression, which were both associated with the cervical lesions, were negatively correlated [r=-0.547(AcH3), r=-0.585(RAR-β2), P<0.05]. Conclusion:Histone acetylation modification is associated with the regulation of RAR-β2 expression. This pro-cess is also likely to participate in the carcinogenesis of cervical carcinoma.
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Objective:To isolate cancer stem cells from cervical carcinoma and to identify their biological characteristics. Methods:Tumor specimens were obtained from 19 cervical cancer patients at stages ⅠA-ⅡB. Primary cells were cultured in tumor sphere medium (TSM) after mechanical dissociation combined with enzymatic digestion. A series of assays were used to identify the characteristics of the sphere forming cells derived from primary culture. Colony formation was observed by limiting dilution method. MTT assay was used to assess proliferation inhibition by paclitaxel and doxorubicin. Cell surface markers were analysed by fluorescence-activated cell sorter (FACS). The expression of stemness-related genes, drug resistance-related genes, and oncogenes were detected by RT-PCR and Western blotting. Tumorigenicity was evaluated by subcutaneous injection of 1×10~5 sphere-forming cells into nude mice. The tumor formation capability was recorded and pathological classification was performed.Results:After 10 to 15 d culture, the formation of non-adherent spheres could be observed in 8 out of 19 primary tumor cells. The formation ratio was increased with the increase in clinical staging. Sphere-forming cells had colony formation capability. Paclitaxel (100 nmol/L) and doxorubicin (100 nmol/L) inhibited the proliferation of these cells by (77.65±6.46)% and (48.00±7.15)%, respectively. The difference was significant (P<0.01). FACS detection results indicated the phenotypes of sphere-forming cells were CD34~-CD105~-CD44~+CK17~+. RT-PCR detection indicated that spheres expressed stemness-related genes (Oct4 and Piwil2), drug resistance gene ABCG2, and oncogenes (c-myc, sox-2 and stat3). Western blotting further indicated stemness-related protein (Oct4 and Piwil2) expression in spheres. Tumors appeared in all animals at 12 weeks after subcutaneous injection of 1×10~5 sphere forming cells and exhibited a high degree of similarity with the primary tumor in cervical cancer patients. Conclusion:Human cervical cancer stem cells were successfully isolated,which provided a useful model for individualized therapy and evaluation of the therapeutic efficacy for cervical cancer patients.
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Objective To investigate the expression and significance of IL-6 and IL-6 receptor (IL-6R)in hysteromyoma.Methods The expression of IL-6 in hysteromyoma and homologous normal myometrium of 20 cases undergoing total hysterectomy were detected by fluorescent quantitative PCR and western-blot,respectively while IL6R was examined by western-blot and RT-PCR.Results The expression of IL-6 protein and mRNA in homologous normal myometrium were obviously higher than that in hysteromyoma[(0.7996 ±0.0359)vs (0.6904 ±0.0414)and (0.0470±0.0071)vs(0.0283±0.0045 ),P＜0.05] ;No difference was observed in IL-6R protein and mRNA expression between leiomyomas and myometrium[(0.7105 ±0.0520)vs (0.6904 ±0.0532)and (0.6644 ±0.0537 )vs (0.6465±0.0501),P＞0.05 ].Conclusion The lower expression of IL-6 in leiomyomas may contribute to the pathogenesis of hysteromyoma to some extent,and there is no relationship between IL-6R and hysteromyoma.
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Aim To observe the c-fos expression in the caudate-putman of 6-OHDA-lesioned rats after Apomorphine intraperitoneal injection and to investigate the inherent mechanism of it. Methods Rats received a unilateral injection of 6-OHDA into SNc. The abnormal rotations induced by Apomorphine (APO) were investigated 1、7、14 d and 21 d after lesion.Meanwhile dopaminergic derogation and c-fos expression were observed with Nissl body staining,eletronicmicroscopic and immunocytochemistry methods. Results The number of dopaminergic neurons decreased gradually in the lesioned site and its eletronicmicroscopic structure was damaged. When over 80% dopaminergic neurons were lossed , contralateral rotations surpassed 7 r?min~(1) and c-fos expressed in the ipsilateral striatum. Conclusion APO induces c-fos expression in the ipsilateral striatum. Intrinsic relations may exist between dopaminergic neurons deletion and c-fos expression