Advances in prostate cancer research models:From transgenic mice to tumor xenografting models

The identification of the origin and molecular characteristics of prostate cancer(PCa)has crucial implications for personalized treatment.The development of effective treatments for PCa has been limited;however,the recent establishment of several transgenicmouse lines and/or xenografting models is better reflecting the disease in vivo.With appropriate models,valuable tools for elucidating the functions of specific genes have gone deep into prostate development and carcinogenesis.In the present review,we summarize a number of important PCa research models established in our laboratories(PSA-Cre-ERT2/PTEN transgenic mouse models,AP-OX model,tissue recombination-xenografting models and PDX models),which represent advances of translational models from transgenic mouse lines to human tumor xenografting.Better understanding of the developments of these models will offer new insights into tumor progression and may help explain the functional significance of genetic variations in PCa.Additionally,this understanding could lead to new modes for curing PCa based on their particular biological phenotypes.

Prediction of Driver Gene Matching in Lung Cancer NOG/PDX Models Based on Artificial Intelligence

Patient-derived tumor xenografts(PDXs)are a powerful tool for drug discovery and screening in cancer.However,current studies have led to little understanding of genotype mismatches in PDXs,leading to massive economic losses.Here,we established PDX models from 53 lung cancer patients with a genotype matching rate of 79.2%(42/53).Furthermore,17 clinicopathological features were examined and input in stepwise logistic regression(LR)models based on the lowest Akaike information criterion(AIC),least absolute shrinkage and selection operator(LASSO)-LR,support vector machine(SVM)recursive feature elimination(SVM-RFE),extreme gradient boosting(XGBoost),gradient boosting and categorical features(Cat Boost),and the synthetic minority oversampling technique(SMOTE).Finally,the performance of all models was evaluated by the accuracy,area under the receiver operating characteristic curve(AUC),and F1 score in 100 testing groups.Two multivariable LR models revealed that age,number of driver gene mutations,epidermal growth factor receptor(EGFR)gene mutations,type of prior chemotherapy,prior tyrosine kinase inhibitor(TKI)therapy,and the source of the sample were powerful predictors.Moreover,Cat Boost(mean accuracy=0.960;mean AUC=0.939;mean F1 score=0.908)and the eight-feature SVM-RFE(mean accuracy=0.950;mean AUC=0.934;mean F1 score=0.903)showed the best performance among the algorithms.Meanwhile,application of the SMOTE improved the predictive capability of most models,except Cat Boost.Based on the SMOTE,the ensemble classifier of single models achieved the highest accuracy(mean=0.975),AUC(mean=0.949),and F1 score(mean=0.938).In conclusion,we established an optimal predictive model to screen lung cancer patients for non-obese diabetic(NOD)/Shi-scid,interleukin-2 receptor(IL-2R)γ^(null)(NOG)/PDX models and offer a general approach for building predictive models.

Acetyl-11-keto-β-boswellic acid inhibits proliferation and induces apoptosis of gastric cancer cells through the phosphatase and tensin homolog/Akt/cyclooxygenase-2 signaling pathway

BACKGROUND Gastric cancer is one of the most common malignant tumors of the digestive system worldwide,posing a serious danger to human health.Cyclooxygenase(COX)-2 plays an important role in the carcinogenesis and progression of gastric cancer.Acetyl-11-keto-β-boswellic acid(AKBA)is a promising drug for cancer therapy,but its effects and mechanism of action on human gastric cancer remain unclear.AIM To evaluate whether the phosphatase and tensin homolog(PTEN)/Akt/COX-2 signaling pathway is involved in the anti-tumor effect of AKBA in gastric cancer.METHODS Human poorly differentiated BGC823 and moderately differentiated SGC7901 gastric cancer cells were routinely cultured in Roswell Park Memorial Institute 1640 medium supplemented with 10%fetal bovine serum and 1%penicillin/streptomycin.Gastric cancer cell proliferation was determined by methyl thiazolyl tetrazolium colorimetric assay.Apoptosis was measured by flow cytometry.Cell migration was assessed using the wound-healing assay.Expression of Bcl-2,Bax,proliferating cell nuclear antigen,PTEN,p-Akt,and COX-2 were detected by Western blot analysis.A xenograft nude mouse model of human gastric cancer was established to evaluate the anti-cancer effect of AKBA RESULTS AKBA significantly inhibited the proliferation of gastric cancer cells in a dose-and time-dependent manner,inhibited migration in a time-dependent manner,and induced apoptosis in a dose-dependent manner in vitro;it also inhibited tumor growth in vivo.AKBA up-regulated the expression of PTEN and Bax,and downregulated the expression of proliferating cell nuclear antigen,Bcl-2,p-Akt,and COX-2 in a dose-dependent manner.The PTEN inhibitor bpv(Hopic)reversed the high expression of PTEN and low expression of p-Akt and COX-2 that were induced by AKBA.The Akt inhibitor MK2206 combined with AKBA downregulated the expression of p-Akt and COX-2,and the combined effect was better than that of AKBA alone.CONCLUSION AKBA inhibits the proliferation and migration and promotes the apoptosis of gastric cancer cells through the PTEN/Akt/COX-2 signaling pathway.

The Inhibitory Effects of Rh-endostatin(YH-16) in Combination with Radiotherapy on Lung Adenocarcinoma A549 in Mice and the Underlying Mechanisms

In order to investigate the inhibitory effects of Endostar(rh-endostatin,YH-16)in combination with radiotherapy on lung adenocarcinoma A549 in mice and the interaction mechanisms of combined therapy,the transplantation tumor models of A549 lung adenocarcinoma were established.When the largest diameter of tumor reached 1.0cm,all nude mice were randomly divided into 4 groups:Endostar group,radiotherapy group,radiotherapy plus Endostar(combined treatment)group,and control group(n=6 in each group).The largest d...

Scoparone inhibits pancreatic cancer through PI3K/Akt signaling pathway

BACKGROUND Pancreatic cancer is a highly malignant tumor of the gastrointestinal system whose emerging resistance to chemotherapy has necessitated the development of novel antitumor treatments.Scoparone,a traditional Chinese medicine monomer with a wide range of pharmacological properties,has attracted considerable attention for its antitumor activity.AIM To explore the potential antitumor effect of scoparone on pancreatic cancer and the possible molecular mechanism of action.METHODS The target genes of scoparone were determined using both the bioinformatics and multiplatform analyses.The effect of scoparone on pancreatic cancer cell proliferation,migration,invasion,cell cycle,and apoptosis was detected in vitro.The expression of hub genes was tested using quantitative reverse transcription polymerase chain reaction(qRT-PCR),and the molecular mechanism was analyzed using Western blot.The in vivo effect of scoparone on pancreatic cancer cell proliferation was detected using a xenograft tumor model in nude mice as well as immunohistochemistry.RESULTS The hub genes involved in the suppression of pancreatic cancer by scoparone were obtained by network bioinformatics analyses using publicly available databases and platforms,including SwissTargetPrediction,STITCH,GeneCards,CTD,STRING,WebGestalt,Cytoscape,and Gepia;AKT1 was confirmed using qRT-PCR to be the hub gene.Cell Counting Kit-8 assay revealed that the viability of Capan-2 and SW1990 cells was significantly reduced by scoparone treatment exhibiting IC50 values of 225.2μmol/L and 209.1μmol/L,respectively.Wound healing and transwell assays showed that scoparone inhibited the migration and invasion of pancreatic cancer cells.Additionally,flow cytometry confirmed that scoparone caused cell cycle arrest and induced apoptosis.Scoparone also increased the expression levels of Bax and cleaved caspase-3,decreased the levels of MMP9 and Bcl-2,and suppressed the phosphorylation of Akt without affecting total PI3K and Akt.Moreover,compared with the control group,xenograft tumors,in the 200μmol/L scoparone treatment group,were smaller in volume and lighter in weight,and the percentages of Ki65-and PCNA-positive cells were decreased.CONCLUSION Our findings indicate that scoparone inhibits pancreatic cancer cell proliferation in vitro and in vivo,inhibits migration and invasion,and induces cycle arrest and apoptosis in vitro through the PI3K/Akt signaling pathway.

Inhibition of self-renewal and differentiation of HT-29 cells-derived cancer stem-like cells by scutellarin via Hedgehog signaling pathway

OBJECTIVE To investigate the inhibitory effect of scutellarin on the self-renewal and differentiation of HT-29 cells-derived cancer stem-like cells(HT-29CSC)in vitro and in vivo,and to explore its mechanism.METHODS The effect of scutellarin on the growth of HT-29CSC was determined by 3D Culture assay.The effect of scutellarin on growth and transformation of HT-29CSC was probed by soft agar colony formation assay.The effect of scutellarin on the differentiation of HT-29CSC was determined by serum induction differentiation assay in vitro.The effects of scutellarin on the expressions of marker gene Lgr5,target gene c-Myc,proliferation gene CK20 and Nanog gene were measured by quantitative real-time RT-PCR.Investigate the effect of scutellarin on the expression of c-Myc,Gli1,and Lgr5 protein by Western blotting.A subcutaneous xenograft model of colon cancer in nude mice was established and administered by intraperitoneal injection.The change of body weight and tumor size of nude mice were observed every two days.Investi⁃gate the effects of scutellarin on the growth of xenograft tumors in nude mice.The expression of CD133,Lgr5,Gli1,Ptch1,c-Myc,Ki67,CK20,Nanog gene in tumors were measured by quantitative real-time RT-PCR.The expression of c-Myc,Gli1,Lgr5,CD133,Ki67 protein were measured by Western blotting.RESULTS Scutellarin can inhibit the growth of HT-29CSC in 3D culture.Compared with the solvent control group,scutellarin can significantly inhibit the growth and transformation and differentiation of HT-29CSC in vitro(P<0.01).The expression levels of marker genes Lgr5,target gene c-Myc,proliferation gene CK20 and Nanog in HT-29CSC were down-regulated by scutellarin.Scutellarin can reduce the expression of c-Myc,Gli1,and Lgr5 protein in HT-29CSC.Scutellarin can inhibit the growth of colon cancer xenografts,lower CD133,Lgr5,Gli1,Ptch1,c-Myc,Ki67,CK20,and Nanog mRNA level of xenograft tumors,reduce the expression of c-Myc,Gli1,Lgr5,CD133,and Ki67 protein of xenograft tumors in nude mice.CONCLUSION Scutellarin,which is the main component of scutellaria barbata,can inhibit the differentiation of HT-29CSC and the mechanism is to inhibit the activity of Hedgehog signaling pathway.

Vascularization ability of glioma stem cells in different three-dimensional microenvironments

Glioblastoma(GBM)is among the most common and aggressive adult central nervous system tumors.One prominent charac-teristic of GBM is the presence of abnormal microvessels.A significant correlation between angiogenesis and prognosis has been observed.Accurately reconstructing this neovascula-rization and tumor microenvironment through personalized in vitro disease models presents a significant challenge.However,it is crucial to develop new anti-angiogenic therapies for GBM.In this study,3D bioprinted glioma stem cell(GSC)-laden hydrogel scaffolds,hybrid Gsc hydrogels and ceil-free hydrogel scaffolds were manufactured to investigate the vas-cularization ability of GsCs in varying 3D microenvironments.

Effect of all-trans retinoic acid on growth of xenograft tumor and its metastasis in nude mice

Objective To study the effect of all trans retinoic acid on growth of xenograft tumor and its metastasis in nude mice Methods Human gastric cancer BGC 823 and MKN 45 cells were inoculated into spleen subcapsule of nude mice, respectively The nude mice were subsequently administered with all trans retinoic acid every other day Food consuming and body weight of nude mice were measured weekly Six weeks later, the nude mice were killed Xenograft tumors in spleen and metastatic tumors in liver were pathologically examined Microvessel density in the tumors was detected immunohistochemically, and serum carcinoembryonic antigen was measured by radioimmunoassay Results After the nude mice were fed with all trans retinoic acid, the growth of splenic tumor and its liver metastasis were inhibited and the metastatic rates decreased by 50% (BGC 823) and 33 3% (MKN 45), respectively The microvessel density in splenic and hepatic tumors reduced by 28 58% and 35 47% (BGC 823), 19 45% and 14 52% (MKN 45), respectively The concentration of carcinoembryonic antigen decreased by 50 24% (BGC 823) and 48 10% (MKN 45) Conclusion All trans retinoic acid may effectively inhibit the growth of xenograft tumor in spleen and its metastasis to liver in nude mice, which can be corroborated by the decrease of carcinoembryonic antigen and microvessel density

Effect of atorvastatin on tumor growth and metastasis in abreast cancer cell xenograft model and its mechanism

This paper aims to evaluate the effects and the possible mechanisms of atorvastatin on tumor growth and metastasis in a xenograft tumor model.Twenty-four female athymic BALB/C mice with MDA-MB-435 xenograft tumors were randomly assigned to three groups:a control group,a low-dose atorvastatin treatment group,and a high-dose atorvastatin treatment group.The mice in the treatment groups began to be administered with atorvas-tatin(10 or 20 mg/kg per day)when the xenograft tumors reached 1 cm in diameter.At the end of the experiment,the tumor volume and weight and the lung metastasis colonies of each mouse were measured.Western blotting was applied to detect phosphorylation of protein kinase B(PKB,Akt),extracellular signal regulated kinase(ERK),c-Jun N-terminal Kinase(JNK),and the expression of cytochrome P450(CYP)subtype CYP2J2.Atorvastatin suppressed xenograft tumor growth and metastasis both in the low-dose and the high-dose treatment groups(P<0.05).Atorvastatin also decreased the phosphorylated Akt(p-Akt)and p-ERK but increased p-JNK expression.However,atorvastatin did not alter the expression of CYP2J2 in tumor tissue.This suggests that atorvastatin has the efficacy of suppressing tumor growth and metastasis in vivo.These effects were not dependent on down-regulation of CYP2J2 expression.

Synergistic control of sex hormones by 17β-HSD type 7: a novel target for estrogen-dependent breast cancer

17β-hydroxysteroid dehydrogenase(17β-HSD)type 1 is known as a critical target to block the final step of estrogen production in estrogen-dependent breast cancer.Recent confirmation of the role of dyhydroxytestosterone(DHT)in counteracting estrogeninduced cell growth prompted us to study the reductive 17β-HSD type 7(17β-HSD7),which activates estrone while markedly inactivatingDHT.The role ofDHTin breast cancer cell proliferation isdemonstratedby its independent suppression of cell growthin the presence of a physiological concentration of estradiol(E2).Moreover,an integral analysis of a large number of clinical samples in Oncomine datasets demonstrated the overexpression of 17β-HSD7 in breast carcinoma.Inhibition of 17β-HSD7 in breast cancer cells resulted in a lower level of E2 and a higher level of DHT,successively induced regulation of cyclinD1,p21,Bcl-2,and Bik,consequently arrested cell cycle in the G0/G1 phase,and triggered apoptosis and auto-downregulation feedback of the enzyme.Such inhibition led to significant shrinkage of xenograft tumors with decreased cancer cell density and reduced 17β-HSD7 expression.Decreased plasma E2 and elevated plasma DHT levels were also found.Thus,the dual functional 17β-HSD7 is proposed as a novel target for estrogen-dependent breast cancer by regulating the balance of E2 andDHT.Thisdemonstrates aconceptual advance on the general belief that the major role of this enzyme is in cholesterol metabolism.

Radiosensitization of human pancreatic cancer by piperlongumine analogues

Radiotherapy is commonly used to treat advanced pancreatic cancers and can improve survival by2 months in combination with gemcitabine.However,prognosis and survival improvement remain unsatisfactory,and effective therapies are urgently needed.Piperlongumine has been demonstrated to have therapeutic potentials against various cancers.In this study,we synthesized a series of piperlongumine derivatives and provided evidence that piperlongumine derivatives could be used as effective radiosensitizers in pancreatic cancer.Two compounds enhanced the radiosensitivity of Panc-1 and SW1990 cells.In a pancreatic bi-flank xenograft tumor model,they significantly inhibited tumor growth.Piperlongumine derivatives could induce reactive oxygen species(ROS)expression and regulate the Keapl-Nrf2 protective pathway with enhancement of radiation-induced DNA damage,G2/M-phase cell cycle arrest,and apoptosis.Collectively,our data offer a proof of concept for the use of piperlongumine derivatives as a novel class of radiosensitizers for the treatment of pancreatic cancer.

Efficacy of the hypoxia-activated prodrug evofosfamide (TH-302) in nasopharyngeal carcinoma in vitro and in vivo

Background:Tumor hypoxia is considered an important factor in metastasis and disease relapse.Evofosfamide is a hypoxia-activated prodrug that selectively targets the hypoxic regions of solid tumors.As hypoxia-inducible factor-1α(HIF-1α)is overexpressed in nasopharyngeal carcinoma(NPC)tissues,we performed the present study to evaluate the efficacy profile of evofosfamide in NPC.Methods:We evaluated the efficacy of evofosfamide as a single agent or combined with cisplatin(DDP)in the NPC cell lines CNE-2,HONE-1 and HNE-1,and in nude mouse xenograft tumor models.Results:Evofosfamide exhibited hypoxia-selective cytotoxicity in NPC cell lines,with 50%inhibition concentration(IC50)values of 8.33±0.75,7.62±0.67,and 0.31±0.07μmol/L under hypoxia in CNE-2,HONE-1 and HNE-1 cells,respectively.The sensitization ranged from ninefold to greater than 300-fold under hypoxia compared with normoxia controls.The combination of evofosfamide with DDP had a synergistic effect on cytotoxicity in the NPC cell lines by combination index values assessment.Cell cycle G2 phase was arrested after treated with 0.05μmol/L evofosfamide under hypoxia.Histone H2AX phosphorylation(γH2AX)(a marker of DNA damage)expression increased while HIF-1αexpression suppressed after evofosfamide treatment under hypoxic conditions.In the HNE-1 NPC xenograft models,evofosfamide exhibited antitumor activity both as a single agent and combined with DDP.Hypoxic regions in xeno-graft tissue were reduced after both evofosfamide monotherapy and combined therapy with DDP.Conclusions:Our results present preclinical evidence for targeting the selective hypoxic portion of NPC by evofosfa-mide as a single agent and combined with DDP and provide rationale for the potential clinical application of evofosfa-mide for the treatment of nasopharyngeal carcinoma.