Development And Research Of Urolithin A As An Anti-nasopharyngeal Carcinoma Drug

Objective: In order to evaluate the effect and possible mechanism of Urolithin A in inhibiting nasopharyngeal carcinoma,we investigated the effect of Urolithin A on the proliferation of nasopharyngeal carcinoma CNE1 and CNE2 cells through in vitro experiments,and observed the effect of urolithin A on nasopharyngeal carcinoma CNE1 and the effect of CNE2 cell migration ability,to explore the changes of ECM receptor interaction signaling pathway,to study the mechanism of urolithin A through affecting the migration of nasopharyngeal carcinoma CNE1 and CNE2 cells to play an anti-nasopharyngeal cancer effect,to observe the effect of Urolithin A on apoptosis of nasopharyngeal carcinoma CNE1 and CNE2 cells.Methods: 1.The effect of urolithin A on the proliferation activity of nasopharyngeal carcinoma CNE1 and CNE2 cells was determined by CCK8 method;2.The changes of cell proliferation after urolithin A acted on nasopharyngeal carcinoma CNE1 and CNE2 cells were observed by plate cloning experiment;3.The effect of urolithin A on the lateral migration ability of nasopharyngeal carcinoma CNE1 and CNE2 cells was tested by scratch healing experiment;4.The effect of urolithin A on the longitudinal migration ability of nasopharyngeal carcinoma CNE1 and CNE2 cells was tested by transwell chamber method;5.Using RNA sequencing technology to detect the potential molecular mechanism of urolithin A effect and nasopharyngeal cancer cells;6.Using bioinformatics to analyze the experimental results obtained by RNA sequencing technology;7.Using Hoechst 33342 and flow cytometry to detect the effect of Uro A on The effect of apoptosis of CNE1 and CNE2 cells;8.The effect of urolithin A on the content of reactive oxygen species in CNE1 and CNE2 cells was detected by reactive oxygen species assay;9.The effect of urolithin A on CNE1 was detected by rhodamine 123 and JC-1 staining solution,CNE2 cell mitochondrial membrane potential;10.Western blot detection of ECM receptor interaction signaling pathway-related proteins,and sequencing showed significantly differentially expressed genes-related proteins and the expression of apoptosis-related proteins.Results: 1.The results of the cytotoxicity test clearly indicated that compared with the blank control group,with the increase of the concentration of urolithin A,its inhibitory effect on the proliferation of nasopharyngeal carcinoma CNE1 and CNE2 cells gradually increased,and the difference was statistically significant(P<0.001);2.The results of plate cloning experiments showed that after 12 days,with the increase of urolithin A concentration,the proliferation ability of nasopharyngeal carcinoma CNE1 and CNE2 cells gradually decreased(P<0.01 or P<0.001);3.The results of cell scratch experiments showed that With the increase of urolithin A concentration,the in vitro migration ability of nasopharyngeal carcinoma CNE1 and CNE2 cells gradually decreased(P<0.01 or P<0.001).After cancer CNE1 and CNE2 cells,the number of penetrating cells was significantly reduced compared with the blank control group,with a statistically significant difference(P<0.001);4.The RNA sequencing results were analyzed by bioinformatics,and the results showed that urolithin A regulates ECM Receptor interaction signaling pathway to the migration of nasopharyngeal carcinoma cells;5.Hoechst 33342 staining and flow cytometry results showed that urolithin A could induce apoptosis of CNE1 and CNE2 cells;6.The reactive oxygen species showed that with the increase of urolithin A concentration,The content of reactive oxygen species in CNE1 and CNE2 cells gradually increased;7.After staining with rhodamine 123 and JC-1,the results showed that with the increase of urolithin A concentration,the mitochondrial membrane potential of CNE1 and CNE2 cells showed a downward trend;8.Molecular immunoblotting showed:(1)Urolithin A has a regulatory effect on the ECM receptor interaction signaling pathway and related proteins of differentially expressed genes;(2)Urolithin A can up-regulate the expression of Bax,c-Caspase-3 and PARP in a concentration-dependent manner.The protein expression of Bcl-2 was down-regulated,and the combined action of urolithin A and NAC could reverse the effect of urolithin A on apoptosis-related proteins.Conclusions: Urolithin A can inhibit the proliferation of nasopharyngeal cancer cells,and inhibit the migration ability of nasopharyngeal cancer cells in vitro by regulating important members in the ECM receptor interaction signaling pathway;it can also be induced by ROS-mediated mitochondrial apoptosis pathway CNE1 and CNE2 cells undergo apoptosis;it provides experimental evidence for the use of urolithin A in the treatment of nasopharyngeal carcinoma.