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Detection of a Circulating MicroRNA Custom Panel in Patients with Metastatic Colorectal Cancer
Published on: March 14, 2019
hsa-miR-29c-3p regulates biological function of colorectal cancer by targeting SPARC
Shitong Zhang1, Jianjun Jin1, Xiaoxiao Tian1
1First Affiliated Hospital of Henan University of Science and Technology, Luoyang City 471003, Henan Province, China.
Abstract:
Colorectal cancer (CRC) is the most common type of behavioral cancers, miRNAs play a critical role in cancer development and progression. In the present study, we downloaded the original data from Gene Expression Omnibus (GEO) and conduct data analysis. has-mir-29c-3p mimic, inhibitor, negative control or si-SPARC (secreted protein acidic, rich in cysteine) were transfected into HCT116 cells, respectively. Quantitative real time PCR (qRT-PCR) was used to measure has-mir-29c-3p and SPARC mRNA expressions, western blot was used to detect ACAA1 (acetyl-CoA acyltransferase 1), ACOX1 (acyl-CoA oxidase 1), COL1A1(collagen, type I, alpha-1), COL1A2 (collagen, type I, alpha-2), COL4A1 (collagen, type IV, alpha-1), COL5A2 (collagen, type V, alpha-2), COL12A1 (collagen, type XII, alpha-1), CPT2 (carnitine palmitoyltransferase 2), ETHE1 (persulfide dioxygenase), HMGCS2 (3-hydroxy-3-methylglutaryl-CoA synthase 2), SPARC, SQRDL (sulfide quinone oxidoreductase), and TST (thiosulfate sulfurtransferase) protein expression. CCK-8 and wound healing assay were employed to verify cell proliferation and migration. The luciferase reporter assay data made sure the target correlation of has-mir-29c-3p and SPARC. Firstly, we found that the expression of has-mir-29c-3p was lower in CRC tissues than in their paired corresponding non-cancerous tissues and there was significant inversed correlation between has-mir-29c-3p and SPARC. Overexpression of has-mir-29c-3p reduced cell proliferation and migration. SPARC was identified as a direct target of has-mir-29c-3p, whose silencing reduced cell proliferation and migration. These data showed that has-mir-29c-3p regulates CRC cell functions through regulating SPARC expression. Taken together, has-mir-29c-3p may function as an oncogenic miRNA targeting SPARC, targeted modulation of has-mir-29c-3p expression may became a potential strategy for the treatment.
Insights
MicroRNA-29c-3p (has-mir-29c-3p) is downregulated in colorectal cancer (CRC) and inhibits tumor growth by targeting SPARC. Modulating has-mir-29c-3p offers a potential CRC treatment strategy.
Area of Science:
- Molecular Oncology
- Cancer Biology
- Gene Regulation
Background:
- Colorectal cancer (CRC) is a prevalent malignancy where microRNAs (miRNAs) significantly influence tumorigenesis.
- The role of specific miRNAs, like has-mir-29c-3p, in CRC development and progression warrants detailed investigation.
Purpose of the Study:
- To investigate the expression and function of has-mir-29c-3p in colorectal cancer.
- To identify the target genes of has-mir-29c-3p and elucidate its regulatory mechanisms in CRC cells.
Main Methods:
- Analysis of Gene Expression Omnibus (GEO) data for miRNA and gene expression.
- Cellular assays including transfection with has-mir-29c-3p mimics/inhibitors, SPARC siRNA, qRT-PCR, Western blot, CCK-8, wound healing, and luciferase reporter assays.
- Detection of protein expression for key molecules involved in cellular metabolism and extracellular matrix.
Main Results:
- has-mir-29c-3p expression was significantly lower in CRC tissues compared to non-cancerous tissues, with an inverse correlation to SPARC.
- Overexpression of has-mir-29c-3p suppressed CRC cell proliferation and migration.
- SPARC was validated as a direct target of has-mir-29c-3p; SPARC silencing also reduced cell proliferation and migration.
Conclusions:
- has-mir-29c-3p acts as a tumor suppressor in colorectal cancer by targeting SPARC.
- The has-mir-29c-3p/SPARC axis regulates CRC cell proliferation and migration.
- Targeted modulation of has-mir-29c-3p presents a potential therapeutic strategy for colorectal cancer.

