1.Screening of differentialy expressed saliva proteins from oral lichen planus patients by two-dimensional fluo-rescence difference gel electrophoresis and mass spectrometry
Tiejun LIU ; Kunshan LI ; Jian LIU ; Yongle QIU ; Jingjing WU ; Xin AN ; Yanzhi XU
Journal of Practical Stomatology 2015;(6):793-796
Objective:To identify differentially expressed saliva proteins of oral lichen planus(OLP)patients by two-dimensional fluo-rescence difference electrophoresis(2-D DIGE)and mass spectrometry(MS).Methods:3 pairs of saliva samples from OLP patients and matched healthy adults were collected.Saliva proteins were separated by 2-D DIGE and identified by liquid chromatography-mass spectrometry(LC-MS).Results:SDS-PAGE examination showed that the electrophoresis bands were clear and protein loss was rare. Protein dots were highly reproducible by 2-D DIGE.In average,the abundance of (31 7 ±71 )saliva protein spots were found in OLP pa-tients.4 highly reproducible spots were identified to be secretory IgA1 ,zincα-2-glycoprotein,salivary amylase and serum albumin by LC-MS and they were at higher level in OLP patients than those in the healthy controls.Conclusion:Secretory IgA1 ,zincα-2-glyco-protein,salivary amylase and serum albumin are highly expressed in the saliva of OLP patients,and may be related to the occurrence and development of oral lichen planus.
2.Experimental study on apoptosis⁃inducing and proliferation inhibition by synergistic delivery of viteporfen/TRAIL by extracellular vesicles in oral squamous cell carcinoma cells
Wenjing Wang ; Kunshan Li ; Tiejun Liu ; Xin Liu ; Yongle Qiu
Acta Universitatis Medicinalis Anhui 2022;57(2):247-253
Objective :
To develop an extracellular vesicles ( EVs) co⁃loading system by synergistically deliver tumor necrosis factor related apoptosis inducing ligand (TRAIL) /verteporfin (VPF) combination to induce apoptosis and inhibit proliferation of OSCC cells .
Methods :
TRAIL/VPF co⁃loaded EVs (MSCT⁃EVs/VPF) was purified and collected though ultracentrifugation and dialysis . The expression of CD63 , CD9 and TRAIL was detected by BCA to confirm the origin of EVs . High performance liquid chromatography (HPLC) was used to detect the drug loading of VPF and draw the release curve in vitro . Cytotoxicity and half inhibitory concentration (IC50 ) were detected by MTT assay . The apoptosis rate was detected by flow cytometry . Finally , Western blot was used to detect the effects of MSCT⁃EVs/VPF on the expression of apoptosis related proteins and Yap in SCC25 cells .
Results:
MSCT⁃EVs/VPF particles were round and well dispersed with a diameter of about 100 nm . The drug loading of the nano system was about ( 15 . 43 ± 0. 44)% , 57. 8% of VPF was released in 10 h and 82. 5% in 45h ; MSCT⁃EVs and VPF could inhibit the growth of SCC25 tumor cells in a dose⁃depe~ndent manner , showing good synergistic effect in the ratio of 10 : 1 - 5 : 1 ( CI < 1 , wt% ) . At the ratio of 100 : 5 100 : 15 ( Mass ratio of MSCT⁃EVs to VPF , wt% ) , the IC50 of MSCT⁃EVs/VPF was significantly lower than that of free MSCT⁃EVs + free VPF group (P < 0. 05) , and showed a more effective inhibition . The high inhibitory effect of MSCT⁃EVs/VPF on squamous cell car cinoma cells was partly due to the regulation of Caspase⁃3 , Bax , BCL⁃2 , mTOR , p ⁃mTOR and YAP .
Conclusion
EVs delivery of a fixed proportion of TRAIL/VPF shows high inhibitory effect on oral squamous cell carcinoma cells , which provides a new idea for the treatment of multidrug⁃resistant tumors .