Journal of Diagnostics Concepts & Practice ›› 2023, Vol. 22 ›› Issue (05): 429-440.doi: 10.16150/j.1671-2870.2023.05.003
• Original articles • Previous Articles Next Articles
QIN Xiaodan, SUN Huiling, PAN Bei, PAN Yuqin, WANG Shukui()
Received:
2023-10-23
Online:
2023-10-25
Published:
2024-03-15
CLC Number:
QIN Xiaodan, SUN Huiling, PAN Bei, PAN Yuqin, WANG Shukui. miR-1229-3p inhibits the malignant progression of colorectal cancer and serves as a potential biomarker[J]. Journal of Diagnostics Concepts & Practice, 2023, 22(05): 429-440.
Figure 1
Downregulation of miR-1229-3p expression in human CRC cells A: Stratified clustering heatmap of differentially expressed miRNAs obtained from RNA sequencing data from CRCs and their corresponding normal tissues in the TCGA database. Downregulated genes were represented in red, upregulated genes in green and the red line represented miR-1229-3p; B: Expression of miR-1229-3p in GSE115513 dataset (normal tissue=649, tumor tissue=750); C: The expression of miR-1229-3p in the tissues of 450 colon cancer patients and 8 normal tissues in the Starbase database; D: qRT-PCR was used to detect the expression level of miR-1229-3p in human colorectal cancer cell line and human normal intestinal epithelial cell line NCM460. *:P<0.05; **:P<0.01; ***:P<0.001.
Figure 2
Bioinformatics analysis of miR-1229-3p biological functional characteristics A: Volcano plot showed the differential genes between the high miR-1229-3p expression group and the low miR-1229-3p expression group; B: GO and KEGG analysis of 725 differential genes; C: GSEA of miR-1229-3p based on GO and KEGG gene sets; D: Correlation analysis of miR-1229-3p with 50 marker pathways by GSVA. *:P<0.05; **:P<0.01; ***:P<0.001.
Figure 3
miR-1229-3p affected CRC cell growth A: Stable transfection SW480 and LoVo cells overexpressed by miR-1229-3p were constructed; B: CCK8 assay was used to explore the effect of miR-1229-3p overexpression on the growth ability of CRC cells; C: Clone formation assay was used to explore the effect of the overexpression of miR-1229-3p on the cloning ability of CRC cells; D: The effect of miR-1229-3p overexpression on the proliferation of CRC cells was explored by EdU assay; E: Tumor growth volume curves of tumor-bearing mice in the NC group and miR-1229-3p OE group. *:P<0.05; **:P<0.01; ***:P<0.001.
Figure 4
miR-1229-3p affected CRC cell migration, invasion, and apoptosis A: Transwell assay was used to explore the effect of miR-1229-3p overexpression on the migration of CRC cells; B: Transwell assay was used to explore the effect of miR-1229-3p overexpression on the invasion ability of CRC cells; C: Construction of miR-1229-3p-knockdown SW480 and LoVo cells; D: Flow cytometry assay was used to explore the effect of miR-1229-3p-knockdown on the apoptosis of CRC cells. **:P<0.01; ***:P<0.001.
Figure 4
SETD7 was a potential target gene for miR-1229-3p A: The target gene of miR-1229-3p was predicted by DIANA, TargetScan, and miRDB databases; B: KEGG enrichment analysis of miR-1229-3p target gene; C: GO analysis of miR-1229-3p target gene; D: Expression of SETD7 in CRC from the Human Protein Atlas database; E:qRT-PCR was used to measure SETD7 in SW480 and LoVo cells when miR-1229-3p was overexpressed. *:P<0.05; ***:P<0.001.
Figure 6
miR-1229-3p from plasma exosomes can distinguish patients with CRC A: Representative image of exosomes from LoVo cells under electron microscopy with a bar of 1.0 μm; B: Particle size analysis of exosomes from LoVo cells; C: Western blot analysis of exosomes positive markers TSG101, CD9 and negative marker Calnexin from LoVo cell exosomes, M is protein Marker, control is HEPG2 cells-CD9/Calnexin/TSG101; D: Expression of miR-1229-3p from plasma exosomes of 60 CRC patients and 60 healthy people; E: Receiver operating characteristic curve of miR-1229-3p on the diagnostic efficacy of CRC. ***:P<0.001.
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