Jiang Ping, Li Yuanli, Yang Xin, Zhou Junhong, Wei Peng
Clin Lab. 2020 Jul 1;66(7). doi: 10.7754/Clin.Lab.2020.191222.
There is currently no clinical biomarker for the early diagnosis of giant cell tumor of bone. Long-chain non-coding RNAs (lncRNAs) have a wide range of important gene regulatory functions and play an important role in the occurrence and development of various malignant tumors. This study mainly screened the differential expression of lncRNAs in patients with giant cell tumor of bone by gene chip technology, verified the biological function. We tried to establish a reference basis for early diagnosis of tumor and prediction of tumor recurrence.
From September 2018 to September 2019, we randomly selected 20 cases of primary giant cell tumor of bone and 20 cases of recurrent giant cell tumor of bone, and 20 cases of bone trauma tissue. First, the differential expression of incRNAs in the bone tissue of the patients was evaluated via utilizing gene chip technology. The gene chip was Human LncRNA Array v 3.0 (8 x 60 K) was completed by Shanghai Kangcheng Biotechnology Co., Ltd. The DAVID online analysis platform was used to analyze the differentially expressed genes by GO and KEGG pathways. The target lncRNAs were screened; real-time quantitative PCR (qRT-PCR) was used to verify the relative expression levels of target lncRNAs in bone tissue and serum of three groups of patients.
Using gene chip technology screening, fold-change (FC) value > 2.0 was used as standard. A total of 1,254 differentially expressed lncRNAs were detected of which 896 were up-regulated and 358 were down-regulated. Additionally, a total of 106 differentially expressed lncRNAs were detected with FC values > 10.0. Among these, 72 lncRNAs were upregulated and 34 lncRNAs were downregulated. We then selected two lncRNAs with up-regulation and down-regulation with the largest fold difference. qRT-PCR analytical results showcased that the expression of AK124776 in bone tissue and serum of patients in the recurrent group was significantly higher than that of the initial group and the normal group. Conversely, for RP11-160A10.2, the expression level in the recurrent group was significantly lower than that in the initial group, and the normal group was the highest; the difference was statistically significant (p < 0.05). Finally, we used the expression levels of AK124776 and RP11-160A10.2 in each group as the diagnostic indicators. According to the receiver operating curve (ROC), the accuracy of AK¬124776 and RP11-160A10.2 in the diagnosis of giant cell tumor of bone (area under the curve), the AUC was 0.865 and 0.877, respectively; the accuracy of predicting recurrence of giant cell tumor of bone was 0.832 and 0.841, respectively.
The early detection of differential expression of lncRNAs in the serum of patients with giant cell tumor of bone is important for the diagnosis of disease and prediction of recurrence. The establishment of stable expression of lncRNAs and simple and easy detection methods are of great value for guiding clinical application.
目前尚无用于骨巨细胞瘤早期诊断的临床生物标志物。长链非编码RNA(lncRNAs)具有广泛重要的基因调控功能,在各种恶性肿瘤的发生发展中起重要作用。本研究主要通过基因芯片技术筛选骨巨细胞瘤患者lncRNAs的差异表达,验证其生物学功能,试图为肿瘤早期诊断及复发预测建立参考依据。
2018年9月至2019年9月,随机选取20例原发性骨巨细胞瘤患者、20例复发性骨巨细胞瘤患者及20例骨创伤组织。首先,利用基因芯片技术评估患者骨组织中lncRNAs的差异表达。基因芯片为上海康成生物工程有限公司完成的人类lncRNA Array v 3.0(8×60 K)。使用DAVID在线分析平台通过GO和KEGG通路分析差异表达基因,筛选出目标lncRNAs;采用实时定量PCR(qRT-PCR)验证三组患者骨组织及血清中目标lncRNAs的相对表达水平。
采用基因芯片技术筛选,以变化倍数(FC)值>2.0为标准,共检测到1254个差异表达的lncRNAs,其中896个上调,358个下调。此外,FC值>10.0时共检测到106个差异表达的lncRNAs,其中72个lncRNAs上调,34个lncRNAs下调。然后选取上调和下调倍数差异最大的两个lncRNAs。qRT-PCR分析结果显示,复发组患者骨组织及血清中AK124776的表达明显高于初发组和正常组。相反,对于RP11-160A10.2,复发组的表达水平明显低于初发组,正常组最高;差异有统计学意义(p<0.05)。最后,将每组中AK124776和RP11-160A10.2的表达水平作为诊断指标。根据受试者工作特征曲线(ROC),AK¬124776和RP11-160A10.2诊断骨巨细胞瘤的准确性(曲线下面积),AUC分别为0.865和0.877;预测骨巨细胞瘤复发的准确性分别为0.832和0.841。
骨巨细胞瘤患者血清中lncRNAs差异表达的早期检测对疾病诊断及复发预测具有重要意义。建立稳定的lncRNAs表达及简便的检测方法对指导临床应用具有重要价值。