Application of miRNA-1268b as a diagnostic marker for lung cancer
A technology of miRNA-1268b and DNA molecules, which can be applied in the direction of DNA/RNA fragments, recombinant DNA technology, microbial measurement/inspection, etc., and can solve problems such as limited application and trauma
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Embodiment 1
[0028] Example 1. Discovery of miRNAs associated with lung cancer cells
[0029] Immortalized human bronchial epithelial cell BEP2D is recorded in the following literature: , Lou Tiezhu, Xiang Xiaoqiong, Wu Dechang. Preliminary study on 238Puα particle-induced transformation of human bronchial epithelial cell BEP2D. Chinese Journal of Lung Cancer, 2000, 3(6): 428 -431.2;
[0030] Tumor cell BERP35T44-1 has high serum resistance, strong anchorage-independent growth ability, fast proliferation speed, and certain migration ability, which is recorded in the following literature: α-particle radiation induces malignant growth of human bronchial epithelial cells Research on the biological characteristics of transformed tumor cells; Hu Yingchun; Academy of Military Medical Sciences, 2003 Master's thesis of the Academy of Military Medical Sciences of the Chinese People's Liberation Army.
[0031] 1. Cell culture: BEP2D cells and BERP35T44-1 cells were cultured in LHC-8 serum-free medi...
Embodiment 2
[0049] Example 2, miRNA-1268b is highly expressed in serum exosomes of A549 tumorigenic nude mice
[0050] 1. Animal model: lung cancer cell A549 ( CCL-185TM) was inoculated into BALB / C athymic nude mice, and tumors formed after 4 weeks. The tumors were detected to be caused by A549 cells by live imaging of small animals, and they were recorded as tumor-forming mice.
[0051] 2. Serum collection: The whole blood of tumor-forming mice and control nude mice was collected by orbital blood collection, and the serum was separated.
[0052] 3. qPCR detection of miRNA-1268b expression
[0053] The RNA reverse transcription of the exosomes of tumor-forming mouse serum and control nude mouse serum was extracted respectively to obtain cDNA as a template. The qPCR detection method was the same as in Example 1. The results are shown in Table 3. It can be seen that, compared with the control nude mice, In contrast, miRNA-1268b was up-regulated in serum of tumorigenic nude mice.
[0054...
Embodiment 3
[0057] Example 3, the application of the expression level of miRNA-1268b in the diagnosis of lung cancer patients
[0058] 1. Extraction of RNA
[0059] Serum source of lung cancer patients: Serum of lung cancer patients were all provided by PLA General Hospital, all of which were diagnosed with lung cancer for the first time, and the patients had not received any treatment including surgery, radiotherapy and chemotherapy. The diagnosis is clear and the pathology is clear.
[0060] Source of normal control serum: All normal control serum was provided by the Physical Examination Center of the General Hospital of the People's Liberation Army, and was obtained from routine physical examination of healthy people.
[0061] The RNA of exosomes in the serum of patients with lung cancer and that of normal controls were extracted respectively.
[0062] 2. qPCR detection
[0063] The serum exosomal RNA of the above lung cancer patients and the normal control serum exosomal RNA were u...
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