Method, chip and application for detecting lung cancer cell sensitivity to x-ray radiation
A lung cancer cell, x-ray technology, applied in biochemical equipment and methods, microbial determination/examination, DNA/RNA fragments, etc., can solve problems such as no relationship between microRNA and tumor radiotherapy sensitivity.
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Embodiment 1
[0064] Conventional molecular biology, microbiology and recombinant DNA techniques within the skill of the art can be used in this example. Such techniques are explained fully in the literature. See, eg, Sambrook, Fritsch & Maniatis, Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press, 1989; DNA Cloning: A Practical Approach, Volumes I and II (ed. D.N. Glover, 1985 ); Oligonucleotide Synthesis (M.J.Gait, ed., 1984); Nucleic Acid Hybridization [B.D.Hames & S.J.Higgins, ed. (1985)]; Transcription and Translation [B.D.Hames & S.J.Higgins, ed. (1984)]; B.Perbal, A Practical Guide to Molecular Cloning (1984); Ausubel, F.M. et al. (eds.); A Current Handbook of Molecular Biology, John Wiley & Sons, Inc., 1994.
[0065] 1. Specimen: select lung cancer cell lines (N-A549) sensitive to radiotherapy (S-A549) and insensitive to radiotherapy, and store them in a -80°C low-temperature refrigerator.
[0066] 2. Total RNA extra...
Embodiment 2
[0110] 1. Construct the overexpression vector of miR-126 (that is, miRNA-126), transfect the SK-MES-1 lung cancer cells, and use the empty load and the mother line as a reference, and use the MTT experiment to detect the changes in the cell proliferation ability after 5Gyx ray irradiation , found that high expression of miR-126 significantly inhibited the proliferation of SK-MES-1 cells (see image 3 ).
[0111] The changes in the radiosensitivity of cells after 0, 2, 4, and 8 Gy x-ray irradiation were detected by colony formation experiments, and it was found that high expression of miR-126 significantly increased the radiosensitivity of SK-MES-1 cells (see Figure 4 ).
[0112] Subsequently, the effect of high expression of miR-126 on radiation-induced apoptosis of SK-MES-1 cells was detected. We found that overexpression of miR-126 could significantly promote the apoptosis of lung cancer cells induced by 4Gy and 8Gy x-ray irradiation (P Figure 5 ).
[0113] The above exp...
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