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| Regulatory effect and diagnostic value of piR-128 on colorectal cancer through DNA methylase 3B |
| GAO Xiaobin WU Xueliang ZHAO Yifeng NIE Shuangfa LIANG Feng ZHANG Yingchun#br# |
| Department of General Surgery, the First Affiliated Hospital of Hebei North University, Hebei Province, Zhangjiakou 075000, China |
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Abstract Objective To detect the regulatory effect and diagnostic value of piR-128 on colorectal cancer through DNA methylase (DNMT) 3B. Methods A total of 54 patients diagnosed with colorectal cancer in the Department of General Surgery, the First Affiliated Hospital of Hebei North University from February 2018 to January 2020 were selected. The expression levels of piR-128 and DNMT1, DNMT3A, and DNMT3B in colorectal cancer tissues and adjacent normal tissues were detected by qRT-PCR. Spearman correlation analysis was used to evaluate the association between piR-128 and DNMT3B, and the relationship between patients with different levels of piR-128 and DNMT3B mRNA expression and their clinical characteristics was further analyzed. Receiver operation characteristic curve was used to evaluate the diagnostic and prognostic value of piR-128 in colorectal cancer. Human colorectal cancer cell line LIM1215 was selected and divided into control group (transfected with blank inhibitor and si-blank), piR-128 inhibitor group (transfected with piR-128 inhibitor), SI-DNMT3B group (transfected with si-DNMT3B) and inhibitor +Vector group (transfected with piR-128 inhibitor and DNMT3B Vector). Western blot and qRT-PCR were used to analyze the mutual regulatory relationship between piR-128 and DNMT3B. Transwell assay, cell scratch assay, and flow cytometry were used to analyze the effects of piR-128 and DNMT3B on the invasion, migration, and apoptosis of colorectal cancer cells. Results The levels of piR-128 and DNMT3B mRNA expression in colorectal cancer tissues were higher than those in adjacent normal tissues, and the differences were statistically significant (P ＜ 0.05). The results of correlation analysis showed that there was a positive correlation between the expression level of piR-128 and DNMT3BmRNA in colorectal cancer tissues (rs ＞ 0, P ＜ 0.05). The levels of piR-128 and DNMT3B mRNA in the piR-128 inhibitor group and inhibitor +Vector group were lower than those in the control group, the levels of DNMT3B mRNA in the si-DNMT3B group were lower than those in the control group, and the levels of DNMT3B mRNA in the inhibitor +Vector group were higher than those in the si-DNMT3B group, the protein levels of DNMT3B in the piR-128 inhibitor group, si-DNMT3B group, and inhibitor +Vector group were lower than those in the control group, and the protein level of DNMT3B in the inhibitor +Vector group were higher than those in the si-DNMT3B group, and the differences were statistically significant (P ＜ 0.05). The absorbance of si-DNMT3B group, piR-128 inhibitor group, and inhibitor +Vector group at 24, 36, 48, and 72 h were lower than those of the control group, and the differences were statistically significant (P ＜ 0.05). The invasion ability and migration rate of cells in the piR-128 inhibitor group, si-DNMT3B group, and inhibitor +Vector group were lower than those in the control group, and the apoptosis rate was higher than that in the control group, the invasion ability and migration rate of cells in the inhibitor +Vector group were higher than those in the si-DNMT3B group and piR-128 inhibitor group, and the apoptosis rate was lower than that in the si-DNMT3B group, the differences were statistically significant (P ＜ 0.05). Conclusion piR-128 participates in the progression of colorectal cancer through DNMT3B.
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