| Background and purpose Colorectal cancer is a common malignant tumor of the digestive system.There are about 1.4 million new cases and about 700,000 deaths worldwide each year,which seriously threatens human health.In order to prevent and treat colorectal cancer,a variety of therapeutic methods are used in practice,but the overall efficacy is still unsatisfactory.Therefore,it is imperative to conduct in-depth research on the pathogenesis of the disease to find effective therapeutic targets.In recent years,non-coding RNAs have been confirmed to be closely related to the occurrence and development of tumors.Piwi protein-interacting RNA(pi RNA)is a newly discovered short-chain non-coding RNA.It has been found that abnormal changes in pi RNA expression can promote gastric cancer,cervical cancer and cancer.The occurrence and development of tumors such as bladder cancer,but there are few studies on pi RNA in colorectal cancer.Our research group found that the expression level of pi R020814 in the serum of colorectal cancer patients was significantly higher than that of healthy people,which has a certain value for the early diagnosis of colorectal cancer.This experiment will detect the expression of pi R020814 in colorectal cancer tissue and study its role in the progression of colorectal cancer.Method1.Real-time quantitative PCR(RT-q PCR)was used to detect the expression level of pi R020814 in colorectal cancer tissues and paired adjacent normal tissues,and to detect the expression of pi R020814 in colon cancer cell lines and colon normal epithelial cell lines The relationship between the clinicopathological parameters of patients and the expression level of pi R020814 was analyzed.2.Stable knockdown and overexpression of pi RNA020814 were constructed using lentiviral vectors,and live cell Incu Cyte S3 dynamic cell imaging analysis,plate clone formation assay,soft agar colony formation assay,Transwell invasion assay,flow cytometry,Nude mouse tumor formation experiments were conducted to study the effect of pi RNA020814 on the malignant behavior of colorectal cancer cells in vivo and in vitro.3.Transcriptome sequencing was used to screen the target genes regμLated by pi R020814 in colorectal cancer cells,and RT-q PCR and Western blot were used to verify the target genes in colorectal cancer tissues and HCT116 and HT29 cells with stable knockdown and overexpression of pi R020814 expression and function.Resμlts1.RT-q PCR resμLts showed that the expression level of pi R020814 in colorectal cancer tissues was significantly higher than that in adjacent normal tissues(P<0.05),and the expression levels in colon cancer cell lines HCT116,HT29 and SW480 were significantly higher than those in colon normal tissues Epithelial cell line NCM460(P<0.05).The expression level of pi RNA020814 in colorectal cancer tissue was correlated with tumor diameter,lymph node metastasis,degree of differentiation,stage,location,etc.,but not with patient age and gender.2.Knockdown of pi RNA020814 strongly inhibited the proliferation,clone formation,anchorage-independent growth and invasion of colon cancer cells,significantly increased the proportion of cells in G0/G1 phase,and significantly decreased the proportion of cells in G2/M phase.It promoted the apoptosis of colon cancer cells and significantlyinhibited the growth of colon cancer transplanted in nude mice;overexpression of pi RNA020814 enhanced the proliferation,clone formation,anchorage-independent growth and invasion of colon cancer cells,and reduced the The proportion of cells in G0/G1 phase and the proportion of cells in G2/M phase were increased,the apoptosis of colon cancer cells was reduced,and the growth of colon cancer xenografts in nude mice was promoted.3.The resμLts of transcriptome sequencing showed that knockdown of pi R020814resμLted in up-regμLation of 61 genes and down-regμLation of 55 genes.GO analysis showed that many differentially expressed genes were closely related to ribosomes.RT-q PCR and Western blot analysis showed that the m RNA expression level of Ribosomal Protein S23(RPS23)in colorectal cancer tissues was significantly positively correlated with the expression level of pi R020814;after knockdown of pi R020814 in colon cancer cells,the m RNA and protein expression levels of RPS23 were significantly decreased;after overexpression of pi R020814 in colon cancer cells,the m RNA and protein expression levels of RPS23 were significantly increased.Rescue experiments showed that interfering with the expression of RPS23 coμLd resist the effects of promoting colon cancer cell proliferation and reducing colon cancer cell apoptosis caused by overexpression of pi R020814.Conclusion pi R020814 promotes the malignant behavior of colon cancer cells by upregμLating the expression of RPS23 and is a potential target for colorectal cancer therapy. |
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