Virtual Library
Start Your Search
Hengyi Chen
Author of
-
+
P2.14 - Targeted Therapy (ID 183)
- Event: WCLC 2019
- Type: Poster Viewing in the Exhibit Hall
- Track: Targeted Therapy
- Presentations: 2
- Now Available
- Moderators:
- Coordinates: 9/09/2019, 10:15 - 18:15, Exhibit Hall
-
+
P2.14-25 - Lorlatinib Induced Protective Autophagy via the AKT–mTOR Pathway in ALK- Rearrangement Lung Cancer Cells (Now Available) (ID 1053)
10:15 - 18:15 | Author(s): Hengyi Chen
- Abstract
Background
Lorlatinib, also known as PF-6463922, is a third-generation anaplastic lymphoma kinase (ALK) tyrosine kinase inhibitor that had been approved clinic treatment with patients who failed previous ALK inhibitors. However, the growth inhibitory effects of Lorlatinib on NSCLC and the underlying mechanism.
Method
The growth inhibitory effects of Lorlatinib were investigated in H3122 and H2228 cell lines. Cell death and proliferation were assessed with MTT and Colony formation assay. Flow cytometry assays was performed to study the cell apoptosis. Cyto-ID® immunofluorescence staining was performed to study the cell autophagy. Signaling transduction was demonstrated with western blot.
Result
We observed that Lorlatinib induces cytotoxicity in H3122 and H2228 cells. trigger autophagy in both H3122 and H2228 cells by increasing the expression of phosphatidylethanolamine‐modified microtubule‐associated protein light‐chain 3 (LC3) and decreasing the expression of p62, still can trigger apoptosis by increasing the expression of B cell lymphoma 2 interacting mediator of cell death (Bim). In the presence of the autophagy blocker (chloroquine) and autophagy enhancer (rapamycin), enhanced the cytotoxicity of Lorlatinib and the Lorlatinib -induced increase in Bim was further augmented. The levels of total and phosphorylated ALK can decrease by Lorlatinib. Lorlatinib inhibited the phosphorylation of AKT and the main autophagy repressor mammalian target of rapamycin (mTOR), pharmacological inhibition of AKT by MK-2206 enhanced Lorlatinib‐induced cell death, and it increased LC3 and Bim level.
Conclusion
We demonstrated that the growth inhibitory effects of Lorlatinib on NSCLC via induced autophagy and apoptosis through AKT/ mTOR signaling pathway, and Pharmacological Intervention of Lorlatinib -induced Autophagy Enhances the Cytotoxicity of Lorlatinib. Our findings provided preliminary data for therapeutic strategies to enhance Lorlatinib efficacy in NSCLC patients.
-
+
P2.14-35 - 2-Deoxy-D-Glucose Sensitizes Non-Small Cell Lung Cancer with EML4-ALK Fusion to Crizotinib via Suppression of HK2 Through AKT/mTOR Passway (ID 749)
10:15 - 18:15 | Author(s): Hengyi Chen
- Abstract
Background
ALK-positive NSCLC cells (ALK+ NSCLC) present high glycolysis. Targeting cancer cell metabolism with the glycolysis inhibitor, 2-deoxyglucose (2DG), is a viable strategy that sensitizes the fist-line ALK-TKI crizotinib; but the effects of combination of 2DG and Crizotinib on ALK+ NSCLC cells and the mechanism of action are unknown.
Method
ALK+ NSCLC cells H2228 and H3122 were incubated with crizotinib with or without 2DG, and subjected to the MTT. H2228 cells was treated with crizotinib or/and 2DG to explore the impact of cell growth and potential mechanism of action by clones formation, Ki67 incorporation assay, small interfering RNA technology, Western blot analysis.
Result
A clear synergistic anti-proliferative interaction between 2DG and crioztinib was observed with a combination index value<1 (CI<1).The combination of crizotinib and 2DG effectively inhibited the clones formation and invasion ability of H2228 cells. The glucose consumption and Lactate production of H2228 cell treated with increasing concentration of 2DG was reduced to accompany a markedly decrease of HK2 expression. ALK-positive NSCLC cells showed a higher level expression of HK2 than ALK-negative NSCLC, down-regulation of HK2 by siRNA, obviously enhanced the ability of crizotinib to suppress proliferation activity. The westblot results displayed a significantly inhibition of AKT/mTOR signaling pathway in H2228 cells treatment with combination of 2DG and crizotinib.
Conclusion
This study demonstrated that 2DG would be a promising drugs to sensitize crizotinib via suppressing HK2 expression to inhibit the activity of AKT/mTOR signaling pathway induced by ALK phosphorylation.
