Renal cell carcinoma miRNA molecular marker and application thereof
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A technology for molecular markers and renal cell carcinoma, applied in the field of miRNA molecular markers for renal cell carcinoma
Active Publication Date: 2021-04-13
GENERAL HOSPITAL OF NUCLEAR IND
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However, which miRNAs can be used as RCC metastas
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Embodiment 1
[0040] Embodiment 1. prepares the test kit (50 times of reaction) of predicting RCC patient's transfer and prognosis
[0041] 1. Trizol: 50ml;
[0042] 2. Chloroform: 20ml;
[0043] 3. Isopropanol: 30ml;
[0044] 4.75% ethanol; 60ml;
[0045] 5. DEPC water: 10ml;
[0046] 6. miRNA reverse transcription specific primer (its sequence is shown in SEQ NO: 1-4): 100ul;
[0047] 7. 200U / ul M-MLV reverse transcriptase: 50ul;
[0048] 8. SYBR qPCR Mix: 500ul;
[0049] 9. 10uM target gene-specific primer (its sequence is shown in SEQ NO:5-10): each 100ul;
[0050] 10. 10uM internal reference gene U6-specific primer (its sequence is shown in SEQ NO: 11-12): 100ul each.
Embodiment 2
[0051] Example 2. Detection and analysis of miRNA molecular markers in tissue samples
[0052] 1. Collect the RCC tissues to be tested, normal paracancerous tissues, and distantly metastatic RCC tissues, wash them with normal saline, and freeze them in liquid nitrogen.
[0053] 2. Tissue RNA extraction: Add liquid nitrogen into the mortar, cut the tissue into pieces and grind it in liquid nitrogen, take 100mg of tissue and add it to 1ml Trizol, and mix well. Leave it at room temperature for 5 minutes, add 200uL / tube of chloroform, shake vigorously and mix for 15 seconds. Centrifuge at 12,000 rpm for 15 minutes. Carefully pipette the upper aqueous phase into a new Ep tube, add an equal volume of isopropanol, invert and mix well, place at room temperature for 5 minutes, and centrifuge at 12,000 rpm for 10 minutes. Carefully discard the supernatant, add 75% ethanol, mix well and centrifuge at 8000 rpm for 8 minutes. Remove the supernatant, and add DEPC water to dissolve the RN...
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Abstract
The invention discloses a renal cell carcinoma miRNA molecular marker which is used as a metastasis marker and is used for diagnosing and/or evaluating prognosis and metastasis risks of renal cell carcinoma, or as a specific molecular target of the renal cell carcinoma which is applied to treatment of the renal cell carcinoma; wherein the miRNA molecular marker comprises one or more of miR-328, miR-502 and miR-504. By detecting the expression level of miR-328, miR-502 and/or miR-504 in the tissue of a renal cell carcinoma patient, whether the renal cell carcinoma patient has metastasis or not and prognosis can be judged, and the accuracy rate reaches 95% or above. Therefore, by detecting miR-328, miR-502 and/or miR-504 and other molecular biomarkers in the tissue of the renal cell carcinoma patient, the disease progression, metastasis and prognosis of the renal cell carcinoma patient can be evaluated, related treatment can be carried out on symptoms in time, the survival time and quality of life of the patient are improved, and great economic value and social benefits are achieved.
Description
technical field [0001] The invention relates to the technical field of biomedicine, more specifically, it relates to renal cell carcinoma miRNA molecular markers and applications thereof. Background technique [0002] Renal cell carcinoma (RCC) is a common cancer worldwide, accounting for about 2-3% of adult malignancies. In 2017, approximately 63,990 new cases of RCC and more than 14,400 kidney cancer-related deaths were reported in the United States. In China, the incidence of RCC is on the rise. In 2014, there were approximately 68,300 new RCC cases and 25,600 RCC-related deaths. Metastatic dissemination is the most important factor affecting the prognosis of RCC. The 5-year survival rate of patients with primary metastases is about 10%, and the 5-year survival rate of patients with non-metastatic and advanced metastases is 70% to 90%. When first diagnosed, nearly 60% of patients were localized cancer, and 20% were distant metastases. The metastatic spread of RCC main...
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