Application of a long non-coding RNA lncLCIR-1 as a molecular marker for lung cancer
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
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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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