The Distribution Of TCM Syndromes In Osteosarcoma And The Study Of Diallyl Trisulfide Inhibiting The Growth Of Osteosarcoma Cells By Regulating Glucose-regulated Protein 78 And Calreticulin

Part 1 The distribution of TCM syndromes in osteosarcomaObjectives: Exploring the distribution of traditional Chinese medicine(TCM)syndromes in osteosarcoma to provide evidence for clinical syndrome differentiation and treatment.Materials and Methods: A questionnaire was made for TCM syndromes,and a total of 30 osteosarcoma patients were collected and analyzed by using SPSS 22.0 Ward system clustering.Results: The ratio of male and female osteosarcoma patients was 3:2;the average age was 19.8 years,and was common in children and young adults.There were four main types of syndrome in osteosarcoma,including qi stagnation and blood stasis syndrome,syndrome of kidney deficiency and damage of essence,phlegm-heat syndrome and syndrome of spleen and kidney yang deficiency,which were related to gender and age.Patients with qi stagnation and blood stasis syndrome were prone to male cases while patients with phlegm-heat syndrome were prone to female cases,and patients with qi stagnation and blood stasis syndrome were younger than those with syndrome of spleen and kidney yang deficiency.Conclusion: There were four main types of syndrome in osteosarcoma,including qi stagnation and blood stasis syndrome,syndrome of kidney deficiency and damage of essence,phlegm-heat syndrome and syndrome of spleen and kidney yang deficiency,which were related to gender and age.Patients with qi stagnation and blood stasis syndrome were prone to male cases while patients with phlegm-heat syndrome were prone to female cases,and patients with qi stagnation and blood stasis syndrome were younger than those with syndrome of spleen and kidney yang deficiency.Part 2 The study of diallyl trisulfide inhibiting the growth of osteosarcoma cells by regulating glucose-regulated protein 78 and calreticulinObjectives: Diallyl trisulfide(DATS)is a natural organic sulfur compound isolated from garlic that has good anticancer activity according to many previous reports.There are many studies pointing out that DATS can downregulate expression of the glucose-regulated protein 78(GRP78)and upregulate expression of calreticulin(CRT),which is associated with poor prognosis and drug resistance in various types of human cancers.However,it remains unknown whether DATS has the same effect on human osteosarcoma cells.This study attempted to clarify the potential molecular mechanisms of the action of DATS in human osteosarcoma Saos-2 cells.Materials and Methods: An inverted phase microscope and immunofluorescent staining was used to observe the morphological changes of Saos-2 cells after being cultured in different concentrations of DATS(0,25,50,and 100 μM)for 24 h,or for four time periods(24,48,72,and 96 h)in the same DATS concentration(50 μM).Quantitative real-time polymerase chain reaction and Western blot were used to detect the expression level of GRP78 or CRT mRNA and proteins in Saos-2 cells.The expression level of GRP78 or CRT was suppressed in Saos-2 cells by utilizing small-interfering RNA,and the cells were subsequently used to study the anti-proliferative effects of DATS treatment.Results: The expression level of GRP78 mRNA and proteins was significantlydownregulated,and the expression level of CRT mRNA and proteins was significantly upregulated due to the increased concentration and effective times of DATS(P<0.05).In addition,there were significant associations between GRP78 or CRT silencing and cell proliferation of DATS treatment(P<0.05).Conclusion: DATS inhibits the growth of human osteosarcoma Saos-2 cells by downregulating the expression of GRP78 and upregulating the expression of CRT.