Application of lncRNA combination in the preparation of products for predicting the prognosis of renal clear cell carcinoma and the sensitivity of molecular targeted drug therapy
A technology for clear cell renal cell carcinoma and molecular targeted drugs, which is applied in the fields of molecular biology and medicine, and can solve the problems that the value of histological prognostic markers in clear cell renal cell carcinoma has not been reported in the literature, and the application research of lncRNA is scarce.
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Embodiment 1
[0056] Embodiment 1: Establish RCClnc4 model
[0057] In previous studies, the inventors firstly used the transcriptome sequencing data of 52 pairs of clear cell renal cell carcinoma and normal kidney tissue (from America cohort from TCGA project (see literature Cancer Genome AtlasResearch Network.Comprehensive molecular characterization of clear cell renalcell carcinoma [ J].Nature,2013,499(7456):43)), 51 differentially expressed lncRNAs were screened out, and unsupervised cluster analysis showed that these lncRNAs could successfully distinguish cancer from normal tissues ( figure 1 ), except for one case of tumor tissue, so we took these 51 lncRNAs as candidate lncRNAs for predicting the prognosis risk of early clear cell renal cell carcinoma (TNM stage I-III). We randomly divided 444 patients with stage I-III clear cell renal cell carcinoma in TCGA into a training group (n=222) and an internal validation group (n=222), and there was no significant difference in clinicopatho...
Embodiment 2
[0059] Example 2: Preliminary verification of the RCClnc4 model
[0060] Subsequently, we selected the median of the RCClnc4 score (–0.323) in the training group as the cutoff value, and divided the patients with clear cell renal cell carcinoma in the training group into a low-risk group and a high-risk group. There was no significant difference in clinicopathological characteristics between the two groups ( p>0.05). Most of the patients who died in the training group were concentrated in the RCClnc4 high-risk group, and the Kaplan-Meier survival curve showed that the overall survival rate of patients with clear cell renal cell carcinoma in the RCClnc4 high-risk group in the training group was lower (p image 3 A). We further applied the above calculation formula and fixed cutoff value to the internal verification group from TCGA and the external verification group from Japan (EGAS00001000509, n=88), and the results showed that the RCClnc4 score can still divide the correspondi...
Embodiment 3
[0062] Embodiment 3: Further verification of RCClnc4 model
[0063] Next, we utilized three independent cohorts from China with long-term follow-up (independent validation group 1 from Changhai Hospital (n=1299), independent validation group 2 from Changzheng Hospital (n=346), independent validation group from Southwest Hospital Validation group 3 (n=224)) included 1869 patients with clear cell renal cell carcinoma to verify the efficacy of RCClnc4 score as a prognostic marker model for clear cell renal cell carcinoma.
[0064] The surgically resected cancer tissue specimens of the above clear cell renal cell carcinoma patients were collected; RNA samples were extracted using Qiagen’s RNeasyPlusMiniKit according to the kit instructions; the extracted RNA was quantitatively detected by NanoDrop2000 spectrometer for the concentration and quality of the extracted total RNA. The extracted RNA samples were frozen and stored in a deep freezer at -80°C. The RNA sample collected abov...
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