Abstract:
Objective To investigate the function and molecular mechanism of 3E10 molecule targeting CD24 in regulating the chemosensitivity of liver cancer HuH-7 cells to cisplatin.
Methods Western blot and flow cytometry were used to verify 3E10 recognition specificity for CD24. Annexin V-FITC/PI double staining was used to detect cell apoptosis. The expression level of drug resistance gene and tumor stem cell gene were analyzed by real-time PCR and Western blot, and the activity of JAK/STAT3 and PI3K/AKT signaling pathways were measured by Western blot.
Results 3E10 could effectively identify CD24 protein and HuH-7 cells. 3E10 significantly enhanced the sensitivity of HuH-7 cells to cisplatin (P < 0.05), and the inhibition rate was increased from (10.3±3.0)% to (34.4±10.8)%. 3E10 significantly decreased the expression level of drug resistance gene ABCB1, ABCB5, ABCC1 and tumor stem gene NANOG, CD24, as well as the phosphorylation level of β-catenin in HuH-7 cells (P < 0.05). 3E10 significantly down-regulated the phosphorylation of STAT3 and AKT(P < 0.05).
Conclusion 3E10 molecule targeting CD24 could enhance the sensitivity of HuH-7 cells to cisplatin through reducing the drug resistance, stemness and JAK/STAT3, PI3K/AKT signaling pathway activity of HuH-7 cells.
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