蛋白激酶D1及其磷酸化位点在皮肤鳞状细胞癌、Bowen病及光线性角化病组织中的表达

摘要/Abstract

摘要： 目的探讨蛋白激酶D1（PKD1）及其磷酸化位点pPKD1?tyr463和pPKD1?ser916在鳞状细胞癌（SCC）、Bowen病和光线性角化病（AK）中的表达及意义。方法收集新鲜SCC、Bowen病、AK及正常皮肤组织各10份，RT?PCR法检测各组样本中PKD1在基因水平的表达，Western印迹法检测各组样本中PKD1及其磷酸化位点在蛋白水平的表达。另收集蜡块组织SCC 50份、Bowen病20份、AK 20份及正常表皮组织10份，免疫组化检测PKD1、pPKD1?tyr463及pPKD1?ser916的表达情况。结果正常皮肤组织、SCC、Bowen病和AK组织中PRKD1 mRNA的表达量分别为0.64 ± 0.09、5.37 ± 1.06、2.69 ± 0.72和2.43 ± 0.46，4组间差异有统计学意义（F = 21.37，P < 0.05），且SCC、Bowen病和AK组织的表达水平均显著高于正常组织（P < 0.05），SCC组织又显著高于AK和Bowen病组织（均P < 0.05），而Bowen病与AK组织的表达量差异无统计学意义（P > 0.05）。PKD1总蛋白及pPKD1?tyr463在SCC和Bowen病组织中主要表达在棘层细胞及异形细胞的细胞质和细胞膜，且阳性表达率均显著高于正常皮肤组和AK组（均P < 0.01）；pPKD1?ser916仅在部分高分化SCC癌巢中少量表达，而低分化鳞癌、AK、Bowen病及正常皮肤组织中均未见表达；SCC组中PKD1阳性表达率随鳞癌病理分级的提高而增加，且PKD1与pPKD1?tyr463的表达呈正相关（rc c = 0.479，P < 0.05）。Western印迹检测结果与免疫组化检测结果大致相符。结论 PKD1及其磷酸化位点Tyr463可能参与复层鳞状上皮来源的皮肤肿瘤的形成和进一步发展分化，在皮肤SCC形成进程中PKD1可能通过Tyr463位点活化而发挥促进作用。

Abstract: Gu Jing, Liu Baoguo, Zhou Meng, Miao Guoying, Lyu Chao, Chai Xiaolei Graduate School of Chengde Medical University, Chengde 067000, Hebei, China （Gu J, Zhou M）; Department of Dermatology, Affiliated Hospital of Hebei University of Engineering, Handan 056002, Hebei, China （Liu BG, Miao GY, Lyu C）; Department of Blood Transfusion, Affiliated Hospital of Hebei University of Engineering, Handan 056002, Hebei, China （Chai XL） Corresponding author: Liu Baoguo, Email: lbg66@163.com 【Abstract】 Objective To measure the of protein kinase D1 （PKD1）, tyr463?phos?phorylaed PKD1 （pPKD1?tyr463） and ser916?phos?phorylaed PKD1 （pPKD1?ser916） in squamous cell carcinoma （SCC）, Bowen′s disease （BD） and actinic keratosis （AK）, and to explore their significance. Methods Fresh tissue samples were resected from lesions of patients with SCC （SCC group）, BD （BD group） and AK （AK group）, as well as from normal skin of healthy human controls （control group）, and each group had a sample size of 10. Real?time RT?PCR was performed to measure the mRNA of protein kinase D1 gene （PRKD1）, and Western blot analysis to determine the protein of PKD1, pPKD1?tyr463 and pPKD1?ser916. In addition, immunohistochemical study was conducted to determine the of PKD1, pPKD1?tyr463 and pPKD1?ser916 in another 50 paraffin?embedded skin samples of SCC, 20 samples of BD, 20 samples of AK and 10 normal skin samples. Results PRKD1 mRNA significantly differed among the control group （0.64 ± 0.09）, SCC group （5.37 ± 1.06）, BD group （2.69 ± 0.72） and AK group （2.43 ± 0.46）（F = 21.37, P < 0.05）, and was significantly higher in the SCC, BD and AK groups than that in the control group （P < 0.05）, as well as in the SCC group than that in the AK and BD groups （both P < 0.05）. However, no significant difference in the PRKD1 mRNA was observed between the BD group and AK group （P > 0.05）. Immunohistochemical study showed that the total PKD1 protein and pPKD1?tyr463 in the SCC and BD groups were mainly expressed in the cytoplasm and cell membrane of spinous layer cells and atypical cells, and their rates were significantly higher than those in the AK group and control group （all P < 0.01）. The pPKD1?ser916 was only slightly expressed in some cancer nests of well?differentiated SCC tissues, but not in poorly?differentiated SCC, AK, BD tissues and normal skin tissues. In the SCC group, the rate of PKD1 increased with the increase of the pathological grade of SCC, and the PKD1 was positively correlated with pPKD1?tyr463 （r c c = 0.479, P < 0.05）. Western blot results were consistent with immunohistochemical findings. Conclusion PKD1 and pPKD1?tyr463 may be involved in the development and differentiation of skin tumors derived from stratified squamous epithelium, and PKD1 may exert promotive effects on the formation of cutaneous SCC by activating the Tyr463 phosphorylation site.

顾静刘保国周萌吕超苗国英柴小磊. 蛋白激酶D1及其磷酸化位点在皮肤鳞状细胞癌、Bowen病及光线性角化病组织中的表达[J]. 中华皮肤科杂志, 2017, 50(4): 247-251.doi:

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