Long non-coding RNA and application thereof
A long-chain non-coding, useful technology, applied in the field of biomedicine, can solve problems such as unclearness and changes in downstream cell functions
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Embodiment 1
[0037] Example 1 Identification of long non-coding RNA and analysis of luciferase activity
[0038] Using the UCSC online genome database analysis, it was found that there were a series of potential long-chain non-coding RNAs downstream of KLF6, as shown in Figure 1 (A), so there may be long-chain non-coding RNAs for feedback regulation of KLF6;
[0039] Based on previous reports, miR-181a has a targeted regulatory effect on KLF6; therefore, the analysis of long-chain non-coding RNA transcripts found that one of the long-chain non-coding RNA transcripts, Lnc00705, has four binding sites for miR-181a point, see Figure 1(B);
[0040] The nucleotide sequence of Lnc00705d is shown in SEQ ID No.1, and the nucleotide sequence of its transcript is shown in SEQ ID No.2.
[0041] SEQ ID No.1 is as follows:
[0042] GCGGAATATACGACTCACTATAGGGAGACCCAAGCTGGCTAGCGTTTAAACTTAAGCTTGCCACCTTTGATGTCTAGAATCAGGGGATCCGAATGTTATTTAATGGTGCCAATGATGTTGGAGGATACCTGTGGGAACCTGGATTAGAAGCATGTGCTGCTCTCAGACCTG...
Embodiment 2
[0048] Example 2 Long non-coding RNA targets miR-181a to inhibit the expression of KLF6
[0049] The mucosal cell line GES-1 was used for routine culture. When the cell density reached 70%, Lnc00705siRNA was transfected with Lipofectamine 2000 (Invitrogen), and the medium was changed after 6 hours of transfection, and the culture was continued for 48 hours for the experiment;
[0050] The nucleotide sequence of Lnc00705siRNA is shown in SEQ ID No.3:
[0051] CUGACCAGCAUAGAUGUUA.
[0052] The detection of mRNA and protein expression levels was carried out by qRT-PCR and western blot respectively, and the results were as follows Figure 3(A)-Figure 3(C) shown;
[0053] Depend on Figure 3(A)-Figure 3(C) It can be seen that after inhibiting the expression of Lnc00705, miR-181a is upregulated, while the expression of KLF6 is down-regulated at both mRNA and protein levels.
Embodiment 3
[0054] Example 3 Effect of long non-coding RNA on proliferation and apoptosis of GES-1 cells
[0055] The effect of MTT method and flow cytometry on the proliferation and apoptosis of GES-1 cells after inhibiting Lnc00705, the results are as follows Figure 4(A)-Figure 4(C) shown;
[0056] Depend on Figure 4(A)-Figure 4(C) It can be seen that, compared with the control, inhibiting Lnc00705 promoted the proliferation of GES-1 cells and inhibited cell apoptosis.
[0057] In summary, the present invention provides a long-chain non-coding RNA and its application. The long-chain non-coding RNA is LncRNA00705, which is located downstream of the KLF6 gene. LncRNA00705 acts as a regulatory factor for diseases caused by miR-181a and KLF6 abnormalities, The medicine for preparing related diseases has broad application prospect and market value.
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