Association of hepatocyte-derived growth factor receptor/caudal type homeobox 2 co-expression with mucosal regeneration in active ulcerative colitis

AIM:To characterize the regeneration-associated stem cell-related phenotype of hepatocyte-derived growth factor receptor(HGFR)-expressing cells in active ulcerative colitis(UC).METHODS:On the whole 38 peripheral blood samples and 38 colonic biopsy samples from 18 patients with histologically proven active UC and 20 healthy control subjects were collected.After preparing tissue microarrays and blood smears HGFR,caudal type homeobox 2(CDX2),prominin-1(CD133) and Musashi-1conventional and double fluorescent immunolabelings were performed.Immunostained samples were digitalized using high-resolution Mirax Desk instrument,and analyzed with the Mirax TMA Module software.For semiquantitative counting of immunopositive lamina propria(LP) cells 5 fields of view were counted at magnification x 200 in each sample core,then mean ± SD were determined.In case of peripheral blood smears,30 fields of view with 100 μm diameter were evaluated in every sample and the number of immunopositive cells(mean ± SD) was determined.Using 337 nm UVA Laser MicroDissection system at least 5000 subepithelial cells from the lamina propria were collected.Gene expression analysis of HGFR,CDX2,CD133,leucine-rich repeat-containing G-protein coupled receptor 5(Lgr5),Musashi-1 and cytokeratin20(CK20) were performed in both laser-microdisscted samples and blood samples by using real time reverse transcription polymerase chain reaction(RT-PCR).RESULTS:By performing conventional and double fluorescent immunolabelings confirmed by RT-PCR,higher number of HGFR(blood:6.7 ± 1.22 vs 38.5 ±3.18;LP:2.25 ± 0.85 vs 9.22 ± 0.65;P < 0.05),CDX2(blood:0 vs 0.94 ± 0.64;LP:0.75 ± 0.55 vs 2.11± 0.75;P < 0.05),CD133(blood:1.1 ± 0.72 vs 8.3± 1.08;LP:11.1 ± 0.85 vs 26.28 ± 1.71;P < 0.05)and Musashi-1(blood and LP:0 vs scattered) positive cells were detected in blood and lamina propria of UC samples as compared to controls.HGFR/CDX2(blood:0 vs 1± 0.59;LP:0.8 ± 0.69 vs 2.06 ± 0.72,P < 0.05)and Musashi-1/CDX2(blood and LP:0 vs scattered) coexpressions were found in blood and lamina propria of UC samples.HGFR/CD133 and CD133/CDX2 coexpressions appeared only in UC lamina propria samples.CDX2,Lgr5 and Musashi-1 expressions in UC blood samples were not accompanied by CK20 mRNA expression.CONCLUSION:In active UC,a portion of circulating HGFR-expressing cells are committed to the epithelial lineage,and may participate in mucosal regeneration by undergoing mesenchymal-to-epithelial transition.

中低位局部进展期直肠癌新辅助同步放化疗前后G蛋白耦联受体蛋白的表达变化及其临床意义

目的观察新辅助放化疗前后中低位局部进展期直肠癌组织中富含亮氨酸重复序列的G蛋白耦联受体(Lgr5)蛋白的表达变化,探讨其对新辅助放化疗疗效的预测作用。方法分析2014年1月至2017年12月间在郑州大学附属肿瘤医院接受新辅助放化疗后行根治性手术的42例直肠癌患者临床资料,其中男26例,女16例,年龄范围为38~75岁(中位数58岁)。采用免疫组织化学法检测活检组织及术中留取的肿瘤组织中的Lgr5蛋白的表达。应用SPSS17.0统计软件分析。Lgr5蛋白阳性表达程度、肿瘤临床特征与术前同步放化疗敏感性的关系均采用χ2检验。结果Lgr5蛋白在新辅助放化疗前直肠癌组织中的阳性表达与T分期(P<0.05)、N分期(P<0.05)、直肠癌新辅助放化疗后肿瘤病理完全缓解(pCR)水平(P<0.01)相关。新辅助放化疗前Lgr5蛋白在直肠癌组织中的阳性表达率为73.80%,新辅助放化疗后即术中留取肿瘤组织中Lgr5蛋白的阳性表达率为28.57%,两者比较阳性表达率下降45.23%。结论Lgr5蛋白在中低位局部进展期直肠癌组织中有较高的阳性表达率,新辅助放化疗后Lgr5蛋白的阳性表达减少;Lgr5蛋白表达与直肠癌浸润程度及淋巴结转移状况密切相关,且其表达与新辅助放化疗后的肿瘤病理缓解程度有关。