Study On The Effects And Mechanism Of Peiminine And Glabridin In Inhibiting Urothelial Bladder Carcinoma Cells

Bladder cancer is the most common malignant tumor,and its incidence ranks first in the genitourinary system in my country.Most(>90%)bladder cancer patients are pathologically classified as urothelial bladder carcinoma(UBC),75%of UBC patients are diagnosed as non-muscle invasive bladder cancer,and the rest are muscle invasive or metastatic bladder cancer.Surgery is the main clinical treatment for UBC,combined with radiotherapy,chemotherapy and immunotherapy.Surgery can generally remove most of the lesions and reduce tumor metastasis and spread.Cisplatin-based chemotherapy have always been the standard regimen for UBC treatment,the complete remission rate of UBC patients receiving chemotherapy was 20%-35%,but it may be accompanied by adverse reactions such as cystitis and urethral stricture,many patients even developed drug resistance.Immunotherapy for UBC included intravesical BCG and PD-1/PD-L1 immune checkpoint inhibitors.BCG intravesical instillation is the only current treatment that reduces the risk of cancer progression to muscle-invasive disease.The approval of immune checkpoint inhibitors has also improved first-line outcomes in cisplatin-intolerant patients,increasing overall survival from less than 10 months to 19 months.However,immunotherapy still has problems such as poor prognosis and low response rate.40%of NMIBC patients failed BCG therapy,only 12%of cancer patients responded to immune checkpoint drugs,and immunotherapy is very expensive.In summary,high recurrence,metastasis and mortality rates remain a major challenge for UBC treatment,which imposed a huge burden on patients’health and economic support.Therefore,there is an expedite require to exploit new and effective therapeutic approaches.In recent years,natural compounds have received extensive attention due to their good anti-cancer activity and low toxicity,and some natural compounds have also been developed as first-line anti-cancer drugs.Peiminine(PEI)is a steroidal alkaloid compound extracted from the bulbs of Bulbus Fritillariae Cirrhosae.It has various pharmacological effects such as cough,asthma,expectorant,anti-bacterial,sedative and anti-inflammatory.Glabridin(GLA)is a flavonoid extracted from licorice,which could protect nerves,resist osteoporosis,regulate energy metabolism,skin whitening,and anti-oxidant.These two natural compounds have been shown to exhibit good anti-tumor activity in various cancers such as colon cancer,lung cancer and oral cancer.However,the effect and molecular mechanism of PEI and GLA on UBC are still unclear.In this study BIU-87 and EJ cells were used to study the anti-tumor activity and anti-tumor mechanism of PEI and GLA.This study explored the effects of PEI and GLA on UBC cell lines in vitro and in vivo,respectively.In vitro experiments showed that BIU-87 and EJ cells were treated with different concentrations of PEI and GLA,the cells activity decreased in a time-and dose-dependent manner.The IC50 of PEI on BIU-87 and EJ cells was 710.3μg/m L and 651.1μg/m L,respectively.The IC50 of GLA against BIU-87 and EJ cells were 6.02μg/m L and 4.36μg/m L,respectively.Both PEI and GLA were able to induce apoptosis in BIU-87(PEI:3.57%vs 8.10%;GLA:4.37%vs 18.18%)and EJ(PEI:8.7%vs 15.3%;GLA:11.4%vs 20.8%)cells,PEI treatment arrested the cell cycle in G1 phase,while GLA induced cell cycle arrest in G2 phase.In cell scratch experiments,PEI and GLA were observed to significantly inhibit BIU-87(PEI:37.09%vs16.32%;GLA:33.16%vs 13.43%)and EJ(PEI:39.19%vs 19.69%;GLA:49.10%vs 34.82%)cell migration.In terms of mechanism,it was found that PEI and GLA up-regulated the expression of BIM,BAK1 and CYCS at the gene and protein levels.EJ cells were selected to establish a tumor-bearing mouse model.Treatment with PEI and GLA significantly inhibited the growth of UBC xenografts in nude mice,The tumor volume(PEI:1775 mm~3vs 689.8 mm~3;GLA:1775 mm~3vs 743.5mm~3)and mass(PEI:1.23 g vs 0.48 g;GLA:1.23 g vs 0.53 g)in the PEI and GLA groups were significantly lower than those in the control group.It was shown that PEI and GLA could inhibit the growth of UBC tumor in vivo.In conclusion,this study demonstrated the effects and molecular mechanisms of PEI and GLA on human UBC cell lines in vitro and in vivo for the first time,providing a new strategy for the prevention and treatment of UBC.