M2丙酮酸激酶、Akt、缺氧诱导因子-1在脉络膜黑色素瘤中的表达及意义

Signification and expression of pyruvate kinase M2, Akt and hypoxia inducible factor-1 in choroidal melanoma

背景研究发现肿瘤型M2丙酮酸激酶（M2-PK）已成为多种肿瘤组织的一种新型标志物，脉络膜黑色素瘤（CM）组织中M2-PK的表达及P13K／蛋白激酶B（Akt）通路、缺氧诱导因子-1（HIF-1）对CM生物学行为的影响有重要的临床意义。目的观察M2一PK在CM组织中的表达情况，研究M2-PK、Akt、HIF-1三者表达之间的关系，探讨其在CN发生发展中的作用及意义。方法收集1992--2012年青岛大学医学院附属医院眼科病理室存档的CM标本30例和眼睑色素痣标本30例，采用免疫组织化学染色法检测M2-PK、Akt、HIF-1在两种组织中的表达情况，并结合临床及病理学资料对M2-PK、Akt、HIF-1与肿瘤细胞生物学行为的影响进行分析。采用，检验对M2-PK、Akt、HIF-1在不同细胞类型的CM中、不同大小CM中及不同巩膜浸润情况CM组织中表达率的差异进行比较，并对M2-PK、Akt、HIF-1在CM与眼睑色素痣中表达率的差异进行比较；采用配对资料的关联性分析评估M2-PK、Akt、HIF-1在CM中表达的关联性。结果M2-PK、Akt在CM中表达的阳性率分别为73％（22／30）和80％（24／30），在眼睑色素痣中的表达阳性率分别为40％（12／30）和47％（14／30），二者在CM中的表达阳性率均明显高于眼睑色素痣，差异均有统计学意义（x2=6．787，P=0．009；x2=7．177，P=0．007）；HIF-1在CM中和在眼睑色素痣中的阳性表达率分别为70％（21／30）和50％（15／30），差异无统计学意义（x2=2．500，P=0．114）。M2-PK在梭形细胞型CM中的阳性表达率为57％（8／14），在类上皮细胞型CM中为87％（7／8），在混合细胞型CM中为87％（7／8），总体差异无统计学意义（x2=3．617，P：0．172）。M2-PK在小CM中的阳性表达率为44％（4／9），中等CM为85％（11／13），大CM为87％（7／8），差异无统计学意义（x2=5．508，P=0．064）。M2-PK在未浸润巩膜和浸润巩膜的CM中的阳性表达率分别为50％（7／14）、94％（15／16），差异有统计学意义（x2=7．308，P=0．007）。在CM中M2-PK与Akt、M2-PK与HIF-1、Akt与HIF-1的阳性表达均有关联（x2=12．315，P=0．000；x2=17．175，P=0．000；x2=10．159，P=0．001），列联系数分别为0．539、0．603、0．503。结论M2-PK在CM细胞的增生、侵袭和转移过程中发挥重要作用，其表达和激活可能与P13K／Akt—HIF-1通路有关。

Background Researches showed that tumor pyruvate kinase M2 （M2-PK） is a novel marker of multiple tumors. Investigating the influence of M2-PK expression as well as hypoxia inducible factor-1 （ HIF-1 ） and PI3K/Akt pathway on the biological behaviour of choroidal melanoma （CM） is of an important significance. Objective The purpose of this study was to investigate the expression of M2-PK,Akt and HIF-1 in CM and discuss their effects in the development of CM. Methods Thirty CM samples and 30 nevus samples were collected in Affiliated Hospital of Qingdao University Medical College. The percentages of M2-PK, Akt and HIF-1 expression in CM and nevus were detected by immunohistochemistry,and their association with elinicopathological parameters were statistically analyzed. The differences of the expressions of M2-PK,Akt and HIF-1 were evaluated by Chi-square test, and the correlations between M2-PK and Akt or HIF-1 were analyzed by correlation analysis of paired data. Results The percentages of positive expression of M2-PK and Akt in CM were 73% （22/30） and 80% （24/30）, respectively,and those in nevus were 40% （12/30） and 47% （14/30） respectively, showing significant differences between the two types of＂ tumors （X2 = 6. 787, P = 0. 009 ;X2 = 7. 177, P = O. 007 ）. An insignificant rate was found in HIF-I expression in CM between the two types of tumors （70% versus 50% ,Xz = 2. 500, P = 0. 114）. The positive expressing percentage of M2-PK was 57% （8/14） in spindle cell type CM,87% （7/8） in epithelioid cell type CM and 87% （7/8） in mixed cell type CM ,with significant difference among them （X2 =3. 617,P=0. 172）. M2-PK was positively expressed in44% （4/9） of small CM,85% （11/13） of moderate CM and 87% （7/8） of big CM, presenting a significant difference among them （X= = 5. 508,P = 0. 064）. An increasingly expressing percentage of M2-PK was seen in sclera invasive CM （94%） compared with sclera uninvasive CM （50%） （X2 =7. 308,P=0. 007）. The correlations were seen in the expressing percentage between M2-PK and Akt （X2 = 12. 315, P = 0. 000） as well as between M2-PK and HIF-! （X2 = 17. 175, P = 0. 000） or between Akt and HIF-1 （X= = 10. 159, P = 0. 001 ）. Conclusions The expression of M2-PK upregulates in CM, indicating that M2-PK plays an important role in the survival,development,invasion and metastasis of CM probably by PI3K/Akt-HIF-1 pathway.