Application of a long non-coding RNA lncLCIR-1 as a molecular marker for lung cancer
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A long-chain non-coding, non-small cell lung cancer technology, applied in the field of molecular biology, can solve the problems of little research on the role of lncRNA, and achieve the effect of inhibiting invasion and metastasis, inhibiting proliferation, and good stability
Active Publication Date: 2022-07-01
SHANDONG RES INST OF TUMOUR PREVENTION TREATMENT
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It has been reported that some lncRNAs can affect the radiosensitivity of tumor cells, but most of them are concentrated in reproductive system and digestive system tumors, and the role of lncRNAs in lung cancer radiotherapy is rarely studied
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Embodiment 1
[0036] Example 1. Expression changes of non-coding RNAs in lung cancer cells after radiotherapy
[0037] The expression changes of non-coding RNAs in lung cancer cells after radiotherapy were detected, and the non-coding RNAs closely related to lung cancer radiotherapy were determined by detecting the relative expression of non-coding RNAs in tissue samples of lung cancer patients.
[0038] 1. The half-lethal radiation dose of lung cancer cells
[0039] The human lung cancer cell lines A549, H460, H1975 and H1299 were recovered and cultured, and cultured to the logarithmic growth phase. The cells were trypsinized and suspended in 1640 medium, and the cells were counted. Take different cell gradients (500, 800, 1000, 1500, 2000 / well) and spread them in a 96-well plate to ensure that the cells have sufficient nutrients and observe the cell growth status. After 1 week of selection, the cell confluence reaches 80-90 % of the number of cells plated. After the number of plated cel...
Embodiment 2
[0053] Example 2. Effects of nucleic acid molecules on lung cancer cells
[0054] Different lung cancer cells were transiently transfected with small interfering RNA and overexpression plasmid vector to change the expression of lncLCIR-1 in the cells, and to explore the effect of lncLCIR-1 on the biological functions of lung cancer cell proliferation, clone formation, cycle and invasion.
[0055] 1. Background expression level of lncLCIR-1 in lung cancer cells
[0056] RT-qPCR was used to detect the background expression level of lncLCIR-1 in A549, H460, H1975, and H1299 cells. RNA was extracted from A549, H460, H1975, and H1299 cells, and RT-qPCR was performed after reverse transcription.
[0057] RT-qPCR reaction system: 5 μL of 2×SYBR Premix Ex Taq (TliRNaseH Plus), 0.04 μL of 50×ROXReference Dye, 0.5 μL of upstream and downstream primers (10 μM), 0.6 μL of cDNA, 3.86 μL of ultrapure water, a total of 10 μL.
[0058] RT-qPCR reaction conditions: 94°C for 15s; 95°C for 5s, ...
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Abstract
The invention relates to the application of a long-chain non-coding RNA lncLCIR-1 as a molecular marker for lung cancer. The present invention finds for the first time that lncLCIR-1 is stably and highly expressed in lung cancer tissue, and the level of lncLCIR-1 decreases after radiotherapy, indicating that lncLCIR-1 is a potential indicator for predicting the efficacy of radiotherapy in lung cancer, and is a new molecular marker of lung cancer. The application of the lncLCIR-1 detection reagent in the present invention in the preparation of a reagent for diagnosing the prognosis of lung cancer patients is mainly to use lncLCIR-1 as a detection marker to prepare a corresponding fluorescence quantitative PCR detection kit for detecting the prognosis of lung cancer patients after radiotherapy . In addition, the siRNA provided by the present invention can inhibit the expression of lncLCIR-1, thereby regulating the cell cycle of lung cancer, causing G1 phase arrest of lung cancer cells, and inhibiting the proliferation of lung cancer cells, which is of great significance in the preparation of drugs for the treatment of lung cancer.
Description
technical field [0001] The invention relates to the application of a long-chain non-coding RNA lncLCIR-1 as a molecular marker of lung cancer, and belongs to the technical field of molecular biology. Background technique [0002] At a time when science and technology are extremely developed, malignant tumors are still a major killer threatening human life and health. Lung cancer is a common malignant tumor in the world, and it is also the tumor with the highest morbidity and mortality in my country. Radiotherapy, as a common and effective treatment method for tumors, plays an important role in the treatment of lung cancer, however, the long-term survival rate of patients still needs to be improved. In addition, there are individual differences in the sensitivity of different patients to radiotherapy, and even some patients develop radiotherapy resistance during the treatment process, and there is no unified molecular marker clinically to predict the efficacy and prognosis of...
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