Application of lncRNA in the diagnosis and treatment of liver cancer
A technology for early liver cancer and liver cancer treatment, applied in the field of biomedicine, can solve the problems of lack of treatment methods for liver cancer, lack of early diagnosis and treatment methods for liver cancer in pathogenesis, and inability to develop therapeutic drugs for liver cancer, etc., to achieve the effect of improving accuracy
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Embodiment 1
[0064] Example 1 Screening of gene markers related to liver cancer
[0065] 1. Sample collection
[0066] The cancer tissues and adjacent tissues of 10 patients with liver cancer were collected, and the patients gave informed consent. All the above specimens were obtained with the consent of the organizational ethics committee.
[0067] 2. Preparation of RNA samples
[0068] Use QIAGEN's tissue RNA extraction kit to extract tissue RNA, and the specific steps are in accordance with the instructions.
[0069] 3. Reverse transcription and labeling
[0070] The mRNA was reverse transcribed into cDNA with the Low RNA Input Linear Amplification Kit, and the experimental group and the control group were labeled with Cy3.
[0071] 4. Hybrid
[0072] The gene chip used Cancheng Bio-Human lncRNA Array, and hybridization was performed according to the steps of the chip instruction manual.
[0073] 5. Data processing
[0074] After hybridization, the chip was scanned with an Agilent...
Embodiment 2
[0077] Example 2QPCR sequencing to verify the differential expression of ENSG00000267751 gene
[0078] 1. Perform large-sample QPCR verification on the differential expression of ENSG00000267751 gene. According to the sample collection method in Example 1, 60 samples of liver cancer tissue and 60 adjacent tissue samples were collected.
[0079] 2. The RNA extraction steps were the same as those in Example 1.
[0080] 3. Reverse transcription:
[0081] A 25 μl reaction system was used, and 1 μg of total RNA was used as template RNA for each sample, and the following components were added to the PCR tubes: DEPC water, 5× reverse transcription buffer, 10 mM dNTP, 0.1 mM DTT, 30 μM Oligo dT, 200 U / μl M-MLV, template RNA. Incubate at 42°C for 1 h, 72°C for 10 min, and centrifuge briefly.
[0082] (3) QPCR amplification test
[0083] Primer Design:
[0084] The primer sequences of ENSG00000267751 gene are:
[0085] Forward primer: 5'-ATGCTACTCTTCCCTTTG-3' (SEQ ID NO.5)
[00...
Embodiment 3
[0098] Embodiment 3 analyzes the expression situation of ENSG00000267751 in TCGA database
[0099] 1. Data collection
[0100] The lncRNA expression profile data of 200 liver cancer tissues and 50 paracancerous tissues were collected from the TCGA database, and the expression levels of ENSG00000267751 in liver cancer tissues and paracancerous tissues were analyzed; box plots were drawn.
[0101] 2. ROC curve analysis
[0102] The receiver operating characteristics of ENSG00000267751 were analyzed using the pROC package in R language, the binomial exact confidence space was calculated, and the ROC curve was drawn.
[0103] 3. Results
[0104] The expression level of ENSG00000267751 is as figure 2 As shown, the expression of ENSG00000267751 was significantly up-regulated in liver cancer tissues compared to the control group.
[0105] The ROC curve of ENSG00000267751 is as follows image 3 As shown, the AUC value of ENSG00000267751 is as high as 0.8785, and it has high spec...
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