Han Rongbo, Hensley Patrick J, Li Jieming, Zhang Yang, Stark Timothy W, Heller Allie, Qian Hai, Shi Junfeng, Liu Zeyi, Huang Jian-An, Jin Tengchuan, Wei Xiaowei, Zhou Binhua P, Wu Yadi, Kyprianou Natasha, Chen Jinfei, Yang Xiuwei H
Department of Oncology, Nanjing First Hospital Nanjing, Jiangsu, P. R. China.
Department of Pharmacology and Nutritional Sciences, Department of Molecular and Cellular Biochemistry, and Markey Cancer Center, University of Kentucky Lexington 40536, KY, USA.
Am J Transl Res. 2020 Apr 15;12(4):1428-1442. eCollection 2020.
Owing to the complexity of interacting molecular networks on the cell surface, integrin-associated tetraspanin CD151 remains controversial regarding its clinical importance and functional impact in prostate cancer. The current study evaluated dynamics and clinical importance of CD151 expression and its function in prostate cancer by IHC analysis of two independent patient cohorts (n=80, 181), bioinformatic interrogation of the TCGA database, and evaluation of gene knockdown effect at the cellular level. Our data showed that aside from high mRNA expression, CD151 was primarily localized to intercellular junctions at the plasma membrane in normal prostate glands or benign tissues, regardless of nature of antibodies used. By contrast, in primary tumors from patients with metastatic disease, CD151 was largely localized in the cytosol. Furthermore, the level of the cell-cell junction-linked CD151 was inversely associated with Gleason grade and tumor stage (P<0.001 for both). The portion of primary tumors expressing junctional CD151 was also three-fold less in the metastatic patient population than its counterpart (P<0.001). In line with these observations, CD151 and its associated α3β1 or α6β4 integrin inversely correlated with androgen receptor (AR) at the mRNA level (Spearman coefficient: -0.44, -0.48 and -0.42) in the TCGA cohort. Expression of these adhesion molecules also correlated with DNA methylation in their promoters (Spearman coefficient: -0.37, -0.71 and -0.82). Combined, these data suggest that CD151 and associated integrins are linked to tumor metastasis through AR and the epigenetic program. Meanwhile, CD151 knockdown in E-cadherin-positive tumor cells led to increased cell proliferation and induction of the epithelial-mesenchymal transition (EMT)-like phenotype. Given the strong RGD-binding integrin dependence of EMT-featured tumor cells, we examined focal adhesion kinase (FAK), their key signaling effector, in the above patient cohorts. In contrast to CD151, FAK exhibited positive correlation with tumor grade and stage as well as AR and p53 inactivation at either mRNA, protein or genomic level. Taken together, our results suggest that CD151 represses prostate cancer by antagonizing cell proliferation, EMT and the signaling of RGD-binding integrins. Since this anti-tumorigenic role is prone to the AR-mediated transcriptional and epigenetic regulation, CD151 and possibly α3β1 and α6β4 integrins are of potential biomarkers for metastatic prostate cancer.
由于细胞表面相互作用的分子网络十分复杂,整合素相关的四跨膜蛋白CD151在前列腺癌中的临床重要性和功能影响仍存在争议。本研究通过对两个独立患者队列(n = 80, 181)进行免疫组化分析、对TCGA数据库进行生物信息学探究以及在细胞水平评估基因敲低效应,来评估CD151在前列腺癌中的表达动态、临床重要性及其功能。我们的数据表明,除了mRNA高表达外,无论使用何种抗体,CD151在正常前列腺腺体或良性组织的质膜上主要定位于细胞间连接。相比之下,在转移性疾病患者的原发性肿瘤中,CD151主要定位于细胞质中。此外,细胞间连接相关的CD151水平与Gleason分级和肿瘤分期呈负相关(两者P均<0.001)。在转移性患者群体中,表达连接性CD151的原发性肿瘤比例也比其对应群体少三倍(P<0.001)。与这些观察结果一致,在TCGA队列中,CD151及其相关的α3β1或α6β4整合素在mRNA水平与雄激素受体(AR)呈负相关(斯皮尔曼系数:-0.44、-0.48和-0.42)。这些黏附分子的表达也与其启动子中的DNA甲基化相关(斯皮尔曼系数:-0.37、-0.71和-0.82)。综合来看,这些数据表明CD151和相关整合素通过AR和表观遗传程序与肿瘤转移相关。同时,在E-钙黏蛋白阳性肿瘤细胞中敲低CD151会导致细胞增殖增加并诱导上皮-间质转化(EMT)样表型。鉴于具有EMT特征的肿瘤细胞对RGD结合整合素的强烈依赖性,我们在上述患者队列中检测了其关键信号效应分子黏着斑激酶(FAK)。与CD151相反,FAK在mRNA、蛋白质或基因组水平与肿瘤分级、分期以及AR和p53失活呈正相关。综上所述,我们的结果表明CD151通过拮抗细胞增殖、EMT和RGD结合整合素的信号传导来抑制前列腺癌。由于这种抗肿瘤作用易于受到AR介导的转录和表观遗传调控,CD151以及可能的α3β1和α6β4整合素是转移性前列腺癌的潜在生物标志物。