Research On The Mechanism Of Compound 42 Targeting KLK7 Regulating The Proliferation Of PANC-1 Cell Line

Objective To explore the chemical properties of Compound 42.To elucidate the specific molecular mechanism of Compound 42 as one of the coumarin derivatives inhibiting proliferation-related signaling pathway in PANC-1 cells after tissue kallikrein 7(KLK7)activity was inhibited,and exert anti-pancreatic cancer cell proliferation Subsequently.The experiment established good foundation for elucidating KLK7-mediated malignant tumor cell development.Methods(1)TCGA and GTEX databases were used to analyze the transcriptional level of KLK7 in pancreatic cancer by Sangerbox bioinformatics analysis method,and TCGA pancreatic cancer(PAAD)database was used to analyze the relationship between KLK7 transcriptional level and survival of pancreatic cancer patients by UCSC Xena bioinformatics analysis tool.(2)The chemical properties of compound 42 were detected by UV spectrophotometer within a certain period of time under the biological environment of p H=7.4.(3)The inhibition ability of compound 42 on the proliferation of PANC-1 cell in 50 hours was detected by RTCA.(4)The effect of compound 42 on the cell cycle and apoptosis of PANC-1 cells was detected at 48 h by flow cytometry.(5)TMT quantitative proteomics and transcriptomics were used to study the effect of compound 42 on the content of key protein on the cell membrane and the pathway related to proliferation of PANC-1 cells,and further to elucidate specific molecular mechanisms of Compound 42 in regulating PANC-1 cells proliferationResults(1)KLK7 gene transcription level increased in pancreatic cancer tissue,and was negatively correlated with survival time of pancreatic cancer patients.(2)Within a certain time,the maximum absorption peak of Compound 42 gradually decreased,but the degradation rate became extremely slow with the increase of time,so Compound 42 has a certain stability.(3)RTCA showed that Compound 42 gradually inhibited the proliferation of PANC-1 cells with time,and it was concentration-dependent.(4)The results of flow cytometry showed that compound 42 could arrest the cell cycle of PANC-1 in G0/G1 phase with the increase of concentration,however,it had no obvious effect on apoptosis.(5).Bioinformatics analysis of proteomics and transcriptomics revealed that 48 hours after Compound 42 treated PANC-1 cells,the content of key proteins in PI3K/AKT/m TOR,Ras/Raf/MEK/ERK,Wnt,and Hippo signaling pathways has changed significantly,in addition,the content of p53,EGFR,FAK and the proteins related to cell cycle were changed significantly,and the corresponding transcription level also changed significantly.Therefore,we speculated that Compound 42 may block the above signal pathways,thereby inhibiting the proliferation of PANC-1 cells.Conclusion Based on our previous research,the experiment further showed that the increase of KLK7 transcription level is closely related to the survival rate of patients with pancreatic cancer.And Compound 42 has certain chemical stability and could inhibit the proliferation of PANC-1 cells in a concentration-dependent manner,arrest the cell cycle in the G0/G1 phase,and the cell morphology changes significantly,but it had no obvious effect on apoptosis.The key proteins in Cell cycle,p53,PI3K/Akt/m TOR,Wnt,Ras/Raf/MEK/ERK and Hippo signaling pathways that regulate cell proliferation changed significantly after compound 42 treated PANC-1 cells,which would provide a meaningful guideline for the study of the detailed molecular mechanism of KLK7 regulating PANC-1 cells.