| Breast cancer is one of the most common malignancies that threaten the h ealth of women around the world.Every year tens of thousands of women los e their lives due to breast cancer,and breast cancer is becoming younger.Am ong them,triple-negative breast cancer has received extensive attention because of its high metastasis,recurrence,and poor prognosis.And triple-negative brea st cancer lacks effective therapeutic targets,and currently there is still no effec tive treatment program in the clinic.Long-chain non-coding RNAs are RNAs o f a type greater than 200 nucleotides in length that were once thought to be b y-products of transcription.At present,there is a growing body of evidence tha t long-chain non-coding RNAs regulate gene expression mainly in terms of epi genetic level,transcription level,and post-transcriptional level.Long-chain nonc oding RNA also plays an important role in the development of tumors and pla ys a role in various types of tumors.This article mainly studied the biological process of long-chain non-coding RNA MIR100 HG regulating triple negative b reast cancer cell proliferation and cycle,and the preliminary exploration of the regulation of alteRNAtive splicing by MIR100 HG.The main findings of this article are as follows:1.Expression and prognosis of MIR100 HG in triple negative breast cancer(1)TCGA data analysis showed that the expression of MIR100 HG in tripl e-negative breast cancer was significantly higher in Lumial A,Lumial B,HER2 positive,and triple negative.(2)In MBA-MB-231,HCC1806,BT549,HCC1937,MCF7,T47 D,SKBR3and other breast cancer cell lines,MIR100 HG is in MBA-MB-231,HCC1806,BT549,and HCC1937 four types of triple negative breast cancer cell lines.T he expression is obviously high.(3)Kaplan-Meier survival analysis Analysis of the expression and prognosi s of MIR100 HG in 360 cases of breast cancer showed that the expression leve l of MIR100 HG increased,but the survival rate decreased.2.The effect of MIR100 HG on cell proliferation(1)Knockdown of MIR100 HG in the MDA-MB-231 cell line significantly decreased cell proliferation.(2)MIR100HG was over-expressed in MDA-MB-231 cell line,and cell pr oliferation was significantly enhanced.3.The effect of MIR100 H on cell cycle(1)MIR100HG was knocked down in the MDA-MB-231 cell line and the cell cycle arrested at the G1 phase.(2)MIR100HG was overexpressed in the MDA-MB-231 cell line and the cell cycle arrested at S phase.4.The mechanism of MIR100HG(1)RNA-seq data analysis showed that MIR100 HG was mainly associated with cell proliferation,cell cycle and other signal pathways.Heatmap cluster analysis revealed that MIR100 HG regulates the expression of p27.(2)Knockdown and over-expression of MIR100 HG in MDA-MB-231 cell l ine detected by RT-PCR,Western blot and other experimental methods found t hat MIR100 HG at the RNA level,protein levels affect the expression of p27.(3)The bioinformatics predictions revealed that there are three binding site s on the MIR100 HG and p27,which may be combined by forming an RNA-D NA triple helix.(4)Knockdown of MIR100 HG in MDA-MB-231 cells simultaneously over expresses MIR100 HG full-length,pCDH-MIR100HG-TFO1,TFO2,TFO3,etc.I t was found that pCDH-MIR100HG-TFO1 can complement p27.MIR100 HG m ainly through the formation of RNA-DNA triple helix with p27,regulate the e xpression of p27,thereby affecting cell proliferation and cycle changes in triple negative breast cancer.(5)MIR100HG interacts with hnRNP A1 in the MDA-MB-231 cell line.(6)Through histological analysis,it was found that MIR100 HG can regula te alteRNAtive splicing of CTNND1.In summary,MIR100 HG mainly regulates the expression of p27 through R NA-DNA triple helix formation with p27,thereby affecting the proliferation an d cell cycle of triple negative breast cancer cells,thereby affecting the progress ion of triple negative breast cancer.In addition,it was found that MIR100 HG can interact with hn RNP A1 interacts to regulate alteRNAtive splicing of CTN ND1. |
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