Font Size: a A A

Anti-tumor Activity And Mechanism Of Novel Myricetin Derivative S2-1-5 On Human Liver Cancer Cell Line SMMC-7721 In Vitro

Posted on:2024-09-24Degree:MasterType:Thesis
Country:ChinaCandidate:H L ZhouFull Text:PDF
GTID:2544307085960109Subject:Medical cell biology and genetics
Abstract/Summary:PDF Full Text Request
Background: Primary Liver Cancer(referred to as "liver cancer")is a kind of malignant tumor with high incidence and fatality rate,which seriously threatens human life and health! Chemotherapy is one of the main treatment methods for patients with advanced liver cancer at present,because of its easy recurrence and drug resistance,the effect of chemotherapy is poor,so it is of great practical significance to design,synthesize and screen more sensitive and efficient new anti-liver cancer drugs.Myricetin is a naturally occurring flavonol,which has a wide range of anti-tumor activities.Myricetin derivatives are compounds obtained by taking myricetin as the lead and chemical modification of its structure.Studies have shown that some myricetin derivatives have good inhibitory activity against different tumor cells.However,at present,there are few studies on the antitumor activity and mechanism of myricetin derivatives.Using myricetin as the matrix,our research team designed and synthesized a new myricetin derivative S2-1-5.During the screening of anti-tumor activity in vitro,we found that the compound had good anti-tumor activity.On this basis,we further explored the antitumor activity pattern and mechanism of the compound against human liver cancer cell line SMMC-7721 in vitro.Objective: Exploring the in vitro antitumor activity pattern and mechanism of the designed and synthesized novel myricetin derivative S2-1-5 on human liver cancer cell line SMMC-7721.Methods: MTT assay preliminarily verified the inhibitory activity of S2-1-5 on human liver cancer cell line SMMC-7721 at different concentrations and time,and selected the best concentration of S2-1-5 as the concentration of S2-1-5 in the followup experiment;CCK-8 experiment and plate clone formation experiment were used to study the effect of S2-1-5 on the proliferation of human liver cancer cell line SMMC-7721;Transwell migration and invasion experiment was used to study the effect of S2-1-5 on the migration and invasion of human liver cancer cell line SMMC-7721;Flow cytometry and Western Blot assay were used to study the effect of S2-1-5 on cell cycle of human liver cancer cell line SMMC-7721;Flow cytometry,JC-1 staining and Western Blot assay were used to study the effect of S2-1-5 on human liver cancer cell line SMMC-7721 apoptosis;Whole transcriptome sequencing was conducted to investigate the key signaling pathways and CYP24A1 gene involved in the anti-liver cancer effect of S2-1-5;RT-q PCR(Reverse transcription fluorescence quantitative polymerase chain reaction)to verify the effect of S2-1-5 on CYP24A1 in key signal Pathway;The expression of CYP24A1 in liver cancer tissues was analyzed using TIMER 2.0 database;The si RNA(small interfering RNA)of CYP24A1 was constructed and transfected into human liver cancer cell line SMMC-7721;The interference efficiency of CYP24A1 was verified by Western Blot experiment,and si RNA-CYP24A1 with the best interference effect was screened for subsequent experiments;CCK-8 experiment and plate clone formation experiment were used to study the effect of interfering CYP24A1 on the proliferation of human liver cancer cell line SMMC-7721;Transwell migration and invasion experiment was used to study the effect of interfering CYP24A1 on the migration and invasion of human liver cancer cell line SMMC-7721;Flow cytometry and Western Blot assay were used to study the effect of interfering CYP24A1 on the cell cycle of human liver cancer cell line SMMC-7721;Flow cytometry,DAPI staining and Western Blot assay were used to study the effect of interfering CYP24A1 on apoptosis of human liver cancer cell line SMMC-7721.Results: 1.S2-1-5 had significant inhibitory activity on human liver cancer cell line SMMC-7721 in a time and concentration dependent manner.The optimal concentration of S2-1-5 was 2.04 μM(IC50 value of S2-1-5 for 72 h).2.S2-1-5 inhibited the proliferation,migration and invasion of human liver cancer cell line SMMC-7721,blocked the transformation of human liver cancer cell line SMMC-7721 from S phase to G2 / M phase,and induced apoptosis of human liver cancer cell line SMMC-7721.3.The key signaling pathway of S2-1-5 action was "steroid biosynthesis",and CYP24A1 was screened as the key gene of S2-1-5 action.4.The expressions of SQLE and SC5 D were significantly up-regulated and CYP24A1 was significantly down-regulated after S2-1-5 treatment,which was consistent with the sequencing results.5.CYP24A1 is highly expressed in Liver hepatocellular carcinoma(LIHC)compared with neighboring normal tissues.6.Compared with the NC group,the expression of CYP24A1 in groups si RNA-CYP24A1-998,si RNA-CYP24A1-1569 and si RNA-CYP24A1-1671 was decreased,and the expression of CYP24A1 in group si RNA-CYP24A1-1569 was the lowest and the interference efficiency was the highest.7.Interference with CYP24A1 inhibited the proliferation,migration and invasion of human liver cancer cell line SMMC-7721,blocked the transformation of human liver cancer cell line SMMC-7721 from S phase to G2/M phase,and induced apoptosis of human liver cancer cell line SMMC-7721.Conclusion: 1.The novel myricetin derivative S2-1-5 synthesized by our research team showed good anti-human liver cancer cell line SMMC-7721 activity in vitro.2.S2-1-5 can inhibit the proliferation,migration and invasion of human liver cancer cell line SMMC-7721,induce cycle arrest and promote apoptosis by inhibiting the expression of CYP24A1,thus playing an anti-tumor role in vitro.
Keywords/Search Tags:Myricetin derivatives, liver cancer, human liver cancer cell line SMMC-7721, CYP24A1
PDF Full Text Request
Related items