No abstract

LHRH‐PE40 was used to promote the proliferation of bone marrow derived cell (BMDC) and improve the antigen‐presenting ability of BMDC as well as the immune function via the CD40 signal pathway. LHRH‐PE40 was also implicated in cancer treatment, targeting a variety of cancer cells that express luteinizing hormone‐releasing hormone receptor (LHRHR). In the present study, the mechanism and efficacy of LHRH‐PE40 were addressed in the following three aspects. Enzyme‐linked immunosorbent assay was performed to confirm the binding specificity of LHRH‐PE40 to LHRHR. The killing effect of LHRH‐PE40 on target cells was mediated by LHRHR, which specifically killed LHRHR‐positive target cells while the minimal cytotoxicity of LHRHR‐negative cells is negligible. Spiegelmers, a molecule mutually exclusive with GnRH and developed by Sven Klussmann and Dr Sven Klussmann of NOXXON Pharmaceuticals in Germany, demonstrated that LHRH‐PE40 maintains a combinatory characteristics of LHRH and LHRHR. In the end, the mechanism of LHRH‐PE40 underlying induction of apoptosis at low concentration and prolonged conditions was firstly demonstrated by the basic method of detecting apoptosis to induce apoptosis. It provided a scientific basis for clinical application of LHRH‐PE40 and laid a foundation for the further study of LHRH‐PE40 on inducing apoptosis of target cells. The target cells herein refer to tumor cells that overexpress LHRHR. This study shows that activated DC can more effectively promote the proliferation of CD4+ T cells, and initially proved that DC carrying anti‐CD40 antibody promoted the immune treatment of the tumor. Combining LHRH‐PE40 with anti‐CD40 DCs achieved substantially improved efficacy in killing tumor cells.

Background: Cancer stem cells, quiescent and drug-resistant, have become a therapeutic target. Low-intensity pulsed ultrasound (LIPUS), a new noninvasive physical device, promotes pluripotent stem cell differentiation and is mainly applied in tissue engineering but rarely in oncotherapy. We explored the effect and mechanism of LIPUS on glioma stem cell (GSC) expulsion from quiescence.Methods: Immunofluorescence staining and flow cytometry were used to detect changes of stem hallmarkers. RT-PCR results showed the gene expression levels of stem-related transcription factors. Brud and RNA-seq were performed for cell cycle analysis. Western blotting showed different expressions of key point proteins. Telomeres damage was found by FISH and IF-FISH. Fluorescence detection was used for mitochondrial membrane potential assay and singlet oxygen detection. Tumor xenograft and immunohistochemical staining were performed to confirm the role of low intensity pulsed ultrasound.Results: We found that LIPUS led to attenuated expression of GSC hallmarks, promoted GSC escape from G0 quiescence, and significantly weakened the stemness-related Wnt and Hh pathways. Next, Interestingly, LIPUS transferred sonomechanical energy into recombinant cytochrome C and B5 proteins in vitro, which converted oxygen molecules into singlet oxygen, triggering a telomere crisis. The results in vivo and in vitro confirmed that LIPUS enhanced GSC sensitivity to temozolomide.Conclusion: These results demonstrated that LIPUS “wakes up” GSCs to improve chemotherapy by transferring energy to cyt families and leading to telomere crisis.