ANTP-SMACN7 fusion peptide alone induced high linear energy transfer irradiation radiosensitization in non-small cell lung cancer cell lines

Objective:The aim of the present study was to investigate the mechanisms responsible for the radiation-sensitizing effect of antennapedia proteins,ANTP-SMACN7,on lung cancer cells treated with accelerated carbon and Fe particle irradiation.Methods:The ANTP-SMACN7 fusion peptide was synthesized and linked to fluorescein isothiocyanate to determine its ability to penetrate cells.A549 and NCI-H460 cells,human non-small cell lung cancer(NSCLC)cell lines,were irradiated with X-ray or high linear energy transfer(LET)irradiation with or without ANTP-SMACN7 treatment.Cellular survival,apoptosis,and protein expression were studied by colony formation assays,flow cytometry,and western blot analyses,respectively.Results:ANTP-SMACN7 fusion proteins entered the cells and promoted A549 and NCI-H460 cell high LET irradiation radiosensitization.High LET irradiation was more efficient for clonogenic cell killing and the induction of apoptosis(P<0.05).Treatment with ANTP-SMACN7 significantly reduced the A549 and NCI-H460 cell clone-forming percentages and increased apoptosis through inhibition of the X-linked inhibitor of apoptosis protein and the activation of caspase-3 and caspase-9.Conclusions:Regarding pharmaceutical radiosensitization,these findings provided a way to improve high-LET clinical radiotherapy for NSCLC patients.

BLM helicase inhibition synergizes with PARP inhibition to improve the radiosensitivity of olaparib resistant non-small cell lung cancer cells by inhibiting homologous recombination repair

Objective:We aimed to investigate the radiosensitizing efficacy of the poly-ADP-ribose polymerase(PARP)inhibitor,olaparib,and the Bloom syndrome protein(BLM)helicase inhibitor,ML216,in non-small cell lung cancer(NSCLC)cells.Methods:Radiosensitization of NSCLC cells was assessed by colony formation and tumor growth assays.Mechanistically,the effects of ML216,olaparib,and radiation on cell and tumor proliferation,DNA damage,cell cycle,apoptosis,homologous recombination(HR)repair,and non-homologous end joining(NHEJ)repair activity were determined.Results:Both olaparib and ML216 enhanced the radiosensitivities of olaparib-sensitive H460 and H1299 cells,which was seen as decreased surviving fractions and Rad51 foci,increased total DNA damage,andγH2AX and 53BP1 foci(P<0.05).The expressions of HR repair proteins were remarkably decreased in olaparib-treated H460 and H1299 cells after irradiation(P<0.05),while olaparib combined with ML216 exerted a synergistic radiosensitization effect on olaparib-resistant A549 cells.In addition to increases of double strand break(DSB)damage and decreases of Rad51 foci,olaparib combined with ML216 also increased pDNA-PKcs(S2056)foci,abrogated G2 cell cycle arrest,and induced apoptosis in A549 lung cancer after irradiation in vitro and in vivo(P<0.05).Moreover,Western blot showed that olaparib combined with ML216 and irradiation inhibited HR repair,promoted NHEJ repair,and inactivated cell cycle checkpoint signals both in vitro and in vivo(P<0.05).Conclusions:Taken together,these results showed the efficacy of PARP and BLM helicase inhibitors for radiosensitizing NSCLC cells,and supported the model that BLM inhibition sensitizes cells to PARP inhibitor-mediated radiosensitization,as well as providing the basis for the potential clinical development of this combination for tumors intrinsically resistant to PARP inhibitors and radiotherapy.

Lipid analysis of three special nervonic acid resources in China

Nervonic acid plays an important role in nutrition and function of the human body.Malania oleifera,Acer truncatum and Xanthoceras sorbifolium are China’s unique woody plant rich in nervonic acid in seed oil.This study aims to investigate the lipid composition of these 3 special resources.Their fatty acids were detected by gas chromatography coupled with flame detector(GC-FID).Triglycerides(TAGs)and phospholipids(PLs)were detected by shotgun-mass spectrometry(shotgun-MS).Results showed that M.oleifera oil presented the highest level of nervonic acid(46.20±0.22%)among the 3 oils.Seeds oil of A.truncatum and X.sorbifolium had 3.53±0.20%and 1.83±0.21%nervonic acid respectively.53 species of TAGs and 15 species of PLs were identified in M.oleifera oil,with PLs content of 499.94±22.34μg/g.In A.truncatum oil,PL and TAG species were twice more than those in M.oleifera oil,and its’content of PLs was 76.27±3.21μg/g.In X.sorbifolium oil,75 TAGs and 34 PLs were detected,with the lowest PLs at 23.84±0.17μg/g.The results demonstrated that these 3 vegetable oils have great potential to become nervonic acid supplements for human health.

Dachaihu decoction inhibits hypernutrition-induced liver metastasis from colorectal cancer by maintaining the gut vascular barrier

Background:Colorectal cancer(CRC)is the third most common malignancy and the second deadliest cancer worldwide.Metastasis to the liver,the most common metastatic site in CRC,is the leading cause of death in patients with CRC.Hyperlipidemia,which is common in patients with CRC,promotes CRC progression and metastasis.Hyperlipidemia is commonly observed in obese patients and is often induced by hypernutrition.The underlying mechanism of hypernutrition-induced hyperlipidemia in promoting CRC liver metastasis remains unclear,and there is an unmet need for effective and low-cost treatments for patients with CRC.Methods:A mouse cecum orthotopic CRC model combined with high-fat diet(HFD)feeding,was established to mimic liver metastasis in CRC in obese patients.The effects of Dachaihu decoction(DCHD),a traditional herbal medicine used to treat inflammation and nonalcoholic fatty liver disease,and of the conventional prescription medicine obeticholic acid(OCA)were evaluated.HFD-induced obesity,hyperlipidemia,and CRC liver metastasis were assessed,along with the histology and pathology of the liver and intestine and the expression of metabolic genes in these tissues.The effects of DCHD and OCA on HFD-induced outcomes were evaluated,and human umbilical vein endothelial cells(HUVECs)treated with bile acids(BAs)and DCHD were used to study the underlying mechanisms in vitro.Results:HFD-mediated obesity and hyperlipidemia promoted CRC metastasis,accompanied by disruption of the gut vascular barrier(GVB)and altered bile acid(BA)metabolism.DCHD decreased HFD-induced hyperlipidemia and liver metastasis in CRC,improving overall survival.Those effects of DCHD were equivalent to or better than those of OCA.DCHD regulated the expression of genes of BA metabolism and tight junctions(TJ)to prevent HFDinduced disruption of the GVB.In HUVECs,DCHD prevented the increases in intracellular Ca2t and accumulation of reactive oxygen species induced by primary conjugated BAs,assisting in the maintenance of redox homeostasis and preventing the downregulation of TJ proteins,thereby maintaining the integrity of the endothelial barrier.Conclusions:The data provide a link between hypernutrition and GVB disruption,which contributes to high liver metastasis in patients with CRC.DCHD may represent a novel therapy in CRC,and targeting abnormal lipid metabolism could be a promising therapeutic strategy for avoiding hypernutrition-associated CRC metastasis.