1.Analysis of differential expression profile of circRNA in pancreatic cancer based on high⁃throughput sequencing technology
Xiaowei Fu ; Yonghao Ouyang ; Le Hong ; Gen Sun ; Wanpeng Xin ; Siqing Yi ; Weidong Xiao
Acta Universitatis Medicinalis Anhui 2023;58(1):101-108
Objective:
To screen the differential expression profile of circular RNA (circRNA) in pancreatic cancer and analyze its potential function.
Methods:
Four pairs of pancreatic cancer tissues and corresponding adjacent pancreatic tissues were selected for high⁃throughput sequencing , and the differentially expressed circRNA was screened according to fold change > 2 and P < 0. 05. qRT⁃PCR was used to detect the expression of 5 randomly selected differentially expressed circRNA in 20 pairs of pancreatic cancer tissue samples. GO and KEGG enrichment analysis of differentially expressed circRNA was performed , and the top 50 circRNA⁃microRNA interaction network was constructed. The relationship between hsa_circ_0046523 and the clinicopathological features of pancreatic cancer was further analyzed , and the effects of hsa_circ_0046523 on the proliferation , migration and invasion of pancreatic cancer cells were observed by functional experiments.
Results:
A total of 17 182 circRNA were predicted by high⁃throughput sequencing , of which 302 circRNA were differentially expressed including 137 circRNA were upregulated and 165 circRNA were downregulated in pancreatic cancer tissues. The qRT⁃PCR results showed that the expression levels of hsa_circ_0046523 , hsa_circ_0004220 and hsa_circ_0000690 in pancreatic cancer tis⁃sues were significantly upregulated (P<0.05),while the expression levels of hsa_ ir_0008676 and hisa _ circ_0004416 were significantly downregulated (P<0.05).which was consistent with the sequencing results.GO analysis indicated that differentially expressed circRNA were mainly involved in substrate-dependent cell migration, protein kinase complex and cytoskeletal protein bindinig.KEGG pathway enrichment analysis showed that differentially expressed circRNA were mainly involved in protein digestion and absorption , ABC transporters , central carbon metabolism in cancer,and glycineserine and threonine metabolism pathways.The expression level of hsa_ circ_0046523 was closely correlated with tumor differentiation (P=0.002),T stage (P=0.006),lymph node metastasis (P=0.011) and TNM stage (P=0.001).Compared with HPDE6-C7 cells the expression of hsa_circ_0046523 significantly inc increased in pancreatic cancer SW1990, Mia PaCa-2BxPC-3 and Capan-2 ceIls (P<0.05). hsa_circ_ 0046523 knockdown significantly inhibited the proliferation, migration and invasion of pancreatic cancer Mia PaCa⁃2 BxPC⁃3 cells (P < 0. 05) .
Conclusion
There are characteristic differentially expressed circRNAs in pancreatic cancer tissues , and these differentially expressed circRNAs may be involved in the occurrence and development of pancreatic cancer.