Preliminary Study On The Mechanism Of NLRP3 Inflammasome And Caspase1 In Inducing Acute Lymphoblastic Leukemia Resistance

OBJECTIVE: To investigate whether overexpression of NLRP3 inflammasome and caspase1 is associated with hepatocarcinoma resistance,and whether its increased expression may cause hepatic tumor resistance to chemotherapy drugs,thereby promoting the progression and recurrence of hematological tumors,whether inhibition of inflammatory activity increases the sensitivity of hematological tumors to chemotherapeutic drugs.Method: 1.Screening of cell lines and chemotherapy drugs The cells were stimulated by LPS and ATP and incubated with different chemotherapeutic drugs,CCK8 was used to detect cell viability changes.2.Can the drug resistance be reversed after inhibiting the activity of caspase1 Before being stimulated by LPS and ATP,the positive cell strain was pretreated with Caspase1 inhibitor,and then incubated with chemotherapy drugs.CCK8 method was used to detect cell proliferation.3.Whether it can reverse the drug resistance after inhibiting the activity of NLRP3 Before being stimulated by LPS and ATP,pre-treatment of positive cell lines with NLRP3 inhibitor,and then incubated with chemotherapy drugs,CCK8 method to detect cell proliferation.4.The expression of NLRP3 and Caspase1 RT-PCR and Western Blot method was used to detect the expression of NLRP3 and Caspase1 at the RNA and protein levels.5.Explore the mechanism of NLRP3 and Caspase1 overexpression Flow cytometry was used to detect the expression levels of reactive oxygen species.6.Inhibition of the expression of reactive oxygen species on the production of NLRP3 and Caspase1 and whether it can reverse the drug resistance Before being stimulated by LPS and ATP,the positive cell strain was pretreated with ROS inhibitor,and incubated with chemotherapeutic drugs.The expression of NLRP3 and Caspase1 and the cell proliferation was detected.7.Detection of clinical patient specimens Bone marrow blood samples were collected from patients with relapsed and refractory hematologic tumors.,and the expression levels of NLRP3 and Caspase1 were detected.Results: 1.The acute lymphoblastic leukemia cell line stimulated by LPS and ATP showed resistance to the cytotoxic effects of adriamycin,idarubicin,and cytaracbine.2.After the addition of Caspase1 and NLRP3 inhibitors,the drug resistance of LPS and ATP-stimulated cells was reversed.3.The expression of Caspase1 and NLRP3 was up-regulated by LPS and ATP.VX-765 could inhibit the expression of Caspase1.MCC950 is able to inhibit the expression of NLRP3 and Caspase1.4.The expression of ROS was increased in cells stimulated by LPS and ATP.5.ROS inhibitor can inhibit the expression of NLRP3 and Caspase1,and it has a certain reversal effect on drug resistance.6.The expression of Caspase1 and NLPR3 in refractory and relapsed B-ALL patients was elevated compared with non-relapsed and refractory patients.Conclusion: 1.Overexpression of NLRP3 inflammasome and Caspase1 can induce the resistance of acute B lymphocytic leukemia to doxorubicin,cytara-cbine and vesevea.2.Inhibition of NLRP3 and Caspase1 activity can reverse the drug resistance.3.LPS and ATP can promote the expression of NLRP3 inflammasome and Caspase1 by inducing the production of reactive oxygen species.4.The expression of NLRP3 and Caspase1 is elevated in patients with relapsed and refractory B-ALL.