New uses for trf-leu-cag
A technology of trf-leu-cag and inhibitor, which is applied in the field of new application of tRF-Leu-CAG, can solve limited problems and achieve the effect of inhibiting NSCLC cell proliferation and cell cycle progression
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Embodiment 1
[0019] Example 1: Detection and screening of tRF expression in NSCLC samples by high-throughput sequencing
[0020] We collected 30 pairs of NSCLC sample information through the SRA database, and performed data analysis by means of high-throughput sequencing.
[0021] According to the data analysis, among the 30 pairs of database samples, the distribution range of the number of Reads of tRF is 17-34nt, and the number of Reads of 27 kinds of tRNA showed an obvious upward trend (FC>1.3, P<0.05), while tRNA-Leu and tRNA-Val accounted for the highest proportion, respectively 37% and 26%.
[0022] Through screening, tRF-Leu-CAG was determined as the research object, while in the database samples, tRNA-Leu-CAG-1( figure 1 A) and tRNA-Leu-CAG-2 ( figure 1 B) The number of reads shows an obvious upward trend in cancer.
[0023] By detecting the qRT-PCR product, it can be determined that the length of tRF-Leu-CAG is 34nt ( figure 2 A), is from tRNA-Leu-CAG-1 ( figure 2 B) and tR...
Embodiment 2
[0024] Example 2: Changes in the expression level of tRF-Leu-CAG in human NSCLC cell lines
[0025] We detected the expression level of tRF-Leu-CAG in human lung cancer cell lines by qRT-PCR.
[0026] The cell lines used in the present invention include: H23 is p53 + / - Non-small cell lung cancer cell line, A549 for Kras + / + Non-small cell lung cancer cell line, H1299 for p53 - / - Non-small cell lung cancer cell lines, SPCA-1 for Asian non-small cell lung cancer cell lines, PC-9 and H1650 for lung adenocarcinoma cell lines, 95-D for human metastatic large cell lung cancer cell lines, 16HBE for lung bronchial epithelial cells Tie.
[0027] In this example, the Trizol reagent was used to lyse and extract the total RNA in the sample, and the One StepPrimeScript miRNA cDNA Synthesis Kit (Code No. D350A) of TaKaRa Company was used for reverse transcription to obtain the cDNA template. The kit can perform Poly(A) tailing reaction on small non-coding RNA in the sample, and at the s...
Embodiment 3
[0031] Example 3: tRF-Leu-CAG inhibitor can inhibit the cell proliferation and cell cycle of NSCLC
[0032] Through the BLAST comparison of the NCBI database, it was found that the sequence of miR-1251-5p had a certain similarity with tRF-Leu-CAG. After miR-1251-5p inhibitor was transfected into H1299, it was found that the expression of tRF-Leu-CAG was significantly down-regulated, so miR-1251-5p inhibitor could be used as tRF-Leu-CAG inhibitor ( Figure 4 A). At the same time, it was found that tRF-Leu-CAG inhibitor can inhibit the cell proliferation of H1299 ( Figure 4 B), while inhibiting G1 / S transition, arresting cells in G1 phase ( Figure 4 C).
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