Numb/Notch signaling pathway modulation enhances human pancreatic cancer cell radiosensitivity.
The present study aims to evaluate whether repression of the Numb/Notch signaling pathway affects the radiosensitivity of human pancreatic cancer cell lines. Different doses of X-rays (0, 2, 3, 4, and 5 Gy) were applied to the PANC-1, SW1990, and MIA PaCa-2 human pancreatic cancer cell lines, and the Numb/Notch pathway inhibitor DAPT was added at different doses (0, 1, 3, and 5 μmol/l). MTT assay, colony formation assay, flow cytometry, scratch assay, and Transwell experiments were performed, and qRT-PCR and Western blot were conducted for the detection of Numb expression. Tumorigenicity assay in nude mice was carried out to verify the influence of blocker of the Numb/Notch signaling pathway on the radiosensitivity of xenograft tumors. The MTT assay, colony formation assay and flow cytometry experiments revealed that proliferation decreased as radiation dose increased. The viability of PANC-1 cells at 5 Gy, SW 1990 cells at 4 Gy and 5 Gy, and MIA PaCa-2 cells at 2-5 Gy was significantly lower than that of non-irradiated cells (all P < 0.05). The migration and invasion assays indicated that the PANC-1 cell line was least radiosensitive, while the MIA PaCa-2 cell line was the most radiosensitive. Numb expression significantly increased with increasing radiation dose, whereas the expression of Hes1, Notch1, and Hes5 significantly decreased compared to non-irradiated cells (P < 0.05). Compared to untreated control cells, DAPT dose dependently increased Numb expression and inhibited Notch1, Hes1, and Hes5 expressions at 2 Gy (P < 0.05). Subcutaneous tumorigenicity assay in nude mice demonstrated that DAPT increased the radiosensitivity of PANC-1, SW 1990, and MIA PaCa-2 cells. These findings suggest that Numb/Notch signaling in pancreatic cancer cells is associated with X-ray radiation and that inhibition of the Numb/Notch signaling pathway can enhance radiosensitivity, suggesting that inhibition of the Numb/Notch signaling pathway may serve as a potential target for clinical improvement of the radiosensitivity of pancreatic cancer.
Department of Gastroenterology, 307th Hospital of PLA, Academy of Military Medical Science, No.8 East Street, Feng Tai District, Beijing, 100071, China.
Yi-Liang
YL
Bi
Department of Gastroenterology, 307th Hospital of PLA, Academy of Military Medical Science, No.8 East Street, Feng Tai District, Beijing, 100071, China.
Min
M
Min
Department of Gastroenterology, 307th Hospital of PLA, Academy of Military Medical Science, No.8 East Street, Feng Tai District, Beijing, 100071, China.
Wei
W
Shen
Department of Gastroenterology, 307th Hospital of PLA, Academy of Military Medical Science, No.8 East Street, Feng Tai District, Beijing, 100071, China. liuyan20160510@163.com.
Yan
Y
Liu
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000528:radiation effects; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects / Q000528:radiation effects; D003959:Diamines / Q000494:pharmacology; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000528:radiation effects*; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D010190:Pancreatic Neoplasms / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology / Q000532:radiotherapy*; D012333:RNA, Messenger / Q000235:genetics; D011836:Radiation Tolerance; D011839:Radiation, Ionizing; D060888:Real-Time Polymerase Chain Reaction; D051880:Receptors, Notch / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects / Q000528:radiation effects*; D013844:Thiazoles / Q000494:pharmacology; D000072056:Transcription Factor HES-1 / Q000235:genetics / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D023041:Xenograft Model Antitumor Assays
C000612000:24-diamino-5-phenylthiazole; D003959:Diamines; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; C113026:NUMB protein, human; D012333:RNA, Messenger; D051880:Receptors, Notch; D013844:Thiazoles; D000072056:Transcription Factor HES-1; C078493:HES1 protein, human
Hes1; MIA; Notch1; Numb/Notch·PANC-1; PaCa-2; Pancreatic cancer; Radiosensitivity; SW 1990
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