Antitumor Effect Of CLT003 In Cervical Cancer Cells

Cervical cancer is the second most common malignant tumor in women worldwide,after breast cancer.Although advances in medical science and technology,surgical techniques continue to improve,continuous improvement of radiotherapy and chemotherapy to save many patients,cervical cancer patients brought immeasurable benefits,making cervical cancer mortality,survival has been greatly improved.However,tumor recurrence and drug resistance are still the difficulties of the treatment of cervical cancer,which is the leading cause of death of cervical cancer patients.Therefore,to find new therapeutic targets and treatment strategies has important clinical significance.Compound 4(CLT003)[(2,6-diisopropyl)-5-amino-1-hydroiso-indole-1,3-dione] invented by Professor Chen Danyang is a phthalocyanine Imine analogues for the treatment of retinal edema.In our preliminary experimental work,we found that CLT003 can inhibit the proliferation of cervical cancer and pancreatic cancer cell lines,so we further designed experiments to explore the anticancer effect and mechanism of CLT003 in cervical cancerObjective:1.The effect of CLT003 on the proliferation and apoptosis of cervical cancer HeLa and C33 A cells in hypoxia and microenvironment was examined.The combination chemotherapy of cisplatin and paclitaxel,which is commonly used in cervical cancer,was performed to observe the effect of CLT003 on the combination therapy2.To observe the CLT003 in cervical cancer HeLa,C33 A cells on glucose metabolism.Methods:1.HeLa and C33 A cells were cultured under hypoxic conditions and the effects of CLT003 monotherapy and CLT003 combined with cisplatin or paclitaxel on cell proliferation were observed by CCK8 and EDU.The expressions of apoptosis-related proteins were detected by Annexin V-PI assay and Western-blot.2.To observe the Effect of CLT003 on Cervical Carcinoma Cell glucose uptake by Glucose Uptake Assay Kit.The mRNA and protein levels of HIF-1α,GLUT1 and LDHA were detected by RT-qPCR and Western-blot.Results:1.1CLT003 alone and combined chemotherapy cisplatin or paclitaxel can effectively inhibit the proliferation of cervical cancer HeLa and C33 A cells,and CLT003 inhibition of tumor cells in a dose-dependent manner,within a certain range(0,2,4,8,16,32,64,128uM).With the increasing concentration of CLT003,the inhibitory effect gradually increased.The IC50 of CLT003 in HeLa and C33 A cells was 11.521 uM and 12.529 uM,respectively.1.2 CLT003 combination chemotherapy cisplatin or paclitaxel significantly enhance the sensitivity of cervical cancer HeLa,C33 A cells to chemotherapeutic drugs,in the same killing effect on cancer cells,can significantly reduce the dose of chemotherapy drugs.(The difference was statistically significant,P <0.05).1.3 Detection of apoptosis showed that with the increase of concentration of CLT003(0,2.5,5,10μM),apoptosis increased significantly(the difference was statistically significant,P <0.05).2.The mRNA and protein levels of HIF-1α,GLUT1,LDHA and so on by RT-PCR and Western-blot showed that CLT003 could down-regulate the expression of HIF-1α,GLUT1 and LDHA from the mRNA level and the protein level Statistical significance,P <0.05);CLT003 was also found to reduce glucose uptake in cervical cancer cells under hypoxic / hypoglycemic conditions by glucose uptake assay(P <0.05).Conclusion:1.CLT003 inhibits the proliferation of cervical cancer HeLa and C33 A cell lines in a dose-dependent manner and promotes the apoptosis of cells.Combined chemotherapy with cisplatin or paclitaxel can improve the sensitivity of cervical cancer cells to chemotherapy drugs.2.CLT003 may exert anti-tumor activity by inhibiting the hypoxic glycolysis of HIF-1α and its downstream target genes GLUT1 and LDHA.