Study On TCM Syndrome Types Of Oral Squamous Cell Carcinoma And Mechanism Of PEITC Effect On Cal-27 Cell Line In Vitro

Objective: With the development of traditional Chinese medicine,the application of traditional Chinese medicine in anti-tumor is becoming more widespread.In clinical,we often use the herbs of cruciferous Chinese medicine(white mustard seed,radish seed,wolfberry fruit,etc.)to treat malignant tumor diseases and their complications.Modern medical research reports that the traditional Chinese medicine monomer compound phenylethyl isothiocyanate(PEITC)is a common biological active compound in some cruciferous Chinese medicines.PEITC is an oxidative agonist that produces reactive oxygen species(ROS)that have an anti-tumor effect.In this study,we first reviewed the records of the clinical diagnosis and treatment of oral squamous cell carcinoma in the literature of traditional Chinese medicine.Then we performed the classification of TCM syndromes in 160 patients with modern oral squamous cell carcinoma and explored the treatment of oral squamous cells with some herbs of cruciferae.The possibility of cancer;Finally,we further experiment with PEITC intervention in oral squamous cell carcinoma cell line Cal-27 cells to explore its regulation of oral squamous cell carcinoma cell proliferation,cell biological characteristics,apoptosis and autophagy Its mechanism of action.Methods:1.Clinical studies: We selected 160 patients with oral squamous cell carcinoma.We counted the general conditions of patients and TCM syndromes.Finally,we used frequency analysis and cluster analysis to study the classification of TCM syndromes in oral squamous cell carcinoma.2.Experimental study: We applied PEITC to intervene in oral squamous cell carcinoma cell line Cal-27 cells,and treated with different concentrations of PEITC in Cal-27 cells to observe the difference in proliferation ability of Cal-27 cells,and to find the best drug for the experiment.concentration.We used fluorescent probes to detect changes in intracellular reactive oxygen species levels in different concentrations of PEITC treated with Cal-27 cells.We further investigated the effects of PEITC on the biological characteristics of oral squamous cell carcinoma using cell scratch assay,cell colony assay,Transwell migration,and invasion assay.We also examined the effect of PEITC on the expression of apoptosis proteins,autophagy proteins,and related signaling pathways in Cal-27 cells by Western Blot.We also used transmission electron microscopy to detect the formation of intracellular autophagosomes.Finally,we used Real-time PCR technology to test the effect of PEITC on the relative expression of apoptosis-related genes and autophagy-related genes.Results: 1.Clinical study results:(1)According to statistics,160 male patients with squamous cell carcinoma were found to have sex,age,and site of disease.The ratio of male to female was: 1.76:1;the mean age of patients was 60.29±11.35 years,and the median was 61 years;the incidence of disease The main parts are the tongue,jaw and gingiva.(2)After analyzing and clustering the symptoms and signs of traditional Chinese medicine,we classified the TCM syndromes of oral squamous cell carcinoma into four categories: Yangxu Hanning(70per,43.75%),Yin deficiency(9per,5.63%).fiery syndrome(18per,11.25%),and qi stagnation(63per,39.38%).2.Experimental results:(1)PEITC can inhibit the proliferation of oral squamous cell carcinoma Cal-27 cells(P<0.05),and it is time-dependent.(2)PEITC inhibited the migration,invasion and clonality of oral squamous cell carcinoma Cal-27 cells(P<0.05).(3)PEITC can increase the production of reactive oxygen species in oral squamous cell carcinoma Cal-27 cells.(4)PEITC highly expressed the intercellular adhesion protein E-cadherin and inhibited the expression of cell migration protein Vimentin through ROS pathway,and the difference was statistically significant(P<0.05).(5)PEITC can induce apoptosis by ROS pathway(P<0.05),and also inhibit Akt activity through Akt signaling pathway,which promotes apoptosis(P<0.05).(6)After PEITC acted on oral squamous cell carcinoma Cal-27 cells,intracellular autophagy bodies were observed by transmission electron microscopy.Western Blot and Real-time PCR results confirmed that PEITC can activate oral squamous cell carcinoma Cal-27 autophagy occurred,and increased autophagy and promoted tumor cell death(P < 0.05).Conclusion:Oral squamous cell carcinoma of TCM syndrome type can be seen,Yang xu Hanning syndrome,Qizhixueyu syndrome and heat syndrome as the main syndrome,Yin deficiency syndrome in the oral squamous cell carcinoma is rare.Some cruciferous Chinese herbs are spicy and warm,and they have the power of wen yang huatan and ruan jian san jie tongluo.Clinical application and medicinal theory inference indicate that some cruciferous Chinese medicines can treat pathogenesis as Yangxu Hanning and Qi zhi xue yu.syndrome neoplastic disease.We performed in vitro cell experiments to confirm that some of the Chinese medicine monomeric compounds PEITC of Cruciferae have the ability to inhibit the proliferation,migration,invasion and clonality of oral squamous cell carcinoma cells.PEITC can promote the apoptosis of oral squamous cell carcinoma cells through the ROS pathway and induce apoptosis through the Akt signaling pathway.PEITC can also promote the autophagy of Cal-27 cells,increase the level of autophagy,and promote tumor cell death.This paper provides clinical research basis for the classification of TCM syndromes in contemporary oral squamous cell carcinoma,and provides in vitro experimental data for the effect of PEITC against oral squamous cell carcinoma.This thesis provides the basic experimental research basis for the application of some cruciferous Chinese medicines in oral squamous cell carcinoma.