Attenuated Salmonella carrying pGRIM-19-si-survivin co-expression plasmid inhibits growth of prostate cancer subcutaneous xenografts in vivo

AIM：To explore the effects of pGRIM-19-si-survivin co-expression plasmid carried by human attenuated Salmonella on prostate cancer subcutaneous xenograft growth in nude mice. METHODS：Prostate cancer xenograft model was established in nude mice. Co-expression plasmids carried by attenuated Salmonella were introduced by intraperitoneal injection. The xenograft volumes were monitored timely. Immunohistochemical staining, RT-PCR and TUNEL assay were applied to investigate the related mechanisms that pGRIM-19-si-survivin inhibited tumor growth in vivo. RESULTS：Compared with psi-survivin and pGRIM-19 carried by attenuated Salmonella (control groups), the tumor volumes were reduced markedly in pGRIM-19-si-survivin plasmid group. The mean shrinkage rates were 2.36 and 3.02 times. pGRIM-19-si-survivin co-expression plasmid carried by attenuated Salmonella inhibited survivin expression but strengthened GRIM-19 expression obviously (P<0.05). The mRNA expression of apoptosis-related proteins such as Bcl-xL, Stat3, cyclin D1 and c-Myc was inhibited, and the vascular endothelial growth factor (VEGF) mRNA and Ki67 protein were also inhibited, but the caspase-3 mRNA expression was up-regulated (P<0.05) with significant cell apoptosis. CONCLUSION： pGRIM-19-si-survivin co-expression plasmid carried by human attenuated Salmonella inhibits the growth of prostate cancer subcutaneous xenografts in nude mice by promoting cell apoptosis and inhibiting prostatic cancer proliferation.     

Knockdown of survivin using siRNA promotes prostatic apoptosis in cancer cell DU145

AIM: To investigate the promoting effects of survivin-siRNA on apoptosis of DU145 cells. METHODS: The DNA template coding siRNA against survivin was synthesized and recombinant plasmid pSi-sur was constructed. The recombinant and the two controls, liposome and pSi-scrambled plasmid, were transfected into DU145 cells. The expression of survivin in mRNA and protein was detected by RT-PCR and Western blotting, respectively. Apoptosis was measured by acridine orange staining, Annexin V-FITC labeled flow cytometry and TUNEL assay.RESULTS: Compared to the liposome control, the levels of survivin mRNA and protein in siRNA group were 0.28±0.07 and 0.34±0.05 (n=3, P<0.05) respectively 72 h after transfection. The apoptotic rate of the cells in pSi-sur group was significantly higher than that in two control groups as showed in acridine orange staining, Annexin V-FITC labeled flow cytometry and TUNEL assay.CONCLUSION: Survivin siRNA significantly inhibits the expression of survivin both in mRNA and protein levels, and induces apoptosis of DU145 cells in vitro.     

Effect of HGF gene transfection on human lymphoma xenografts in nude mice

AIM：To explore the promotion effect of hepatocyte growth factor (HGF) gene transfection on human lymphoma xenografts in nude mice. METHODS：The model of human lymphoma xenograft in nude mice was established by transplantation of Raji cells, which were transfected with recombinant plasmid pVITRO2-HGF harboring the HGF gene. The body weight of the nude mice and the tumor size were dynamically monitored and the tumor tissues were obtained after 8 weeks. Additionally, the methods of terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) and immunohistochemistry were used to detect the apoptotic index (AI) and microvessel density (MVD). RESULTS：The success rate of the human lymphoma xenografts in nude mice was 96.7%. The tumor volume in HGF transfection group was significantly greater than that in HGF transfection+VP-16 group and control groups (non-transfection group and empty vector group). The tumor volume in HGF transfection+VP-16 group was also bigger than that in control groups. No difference of the tumor volume between non-transfection group and empty vector group was observed. AI in HGF transfection group was substantially lower than that in control groups. AI in HGF transfection+VP-16 group showed a little higher than that in HGF transfection group, yet was still lower than that in control groups. MVD in HGF transfection group was extraordinary higher than that in control groups, but decreased after VP-16 induction (P<0.01), which was still higher than that in control groups. CONCLUSION：HGF gene transfection significantly promotes the growth of human lymphoma xenografts in nude mice and substantially inhibits the apoptosis presumably owing to promoting tumor angiogenesis and inhibiting tumor cell apoptosis.     

Effect of specific hTERT RNA interference on biological characteristics of colon carcinoma in vivo and in vitro

AIM: To investigate the effect of specific hTERT RNA interference on biological characteristics of colon carcinoma in vivo and in vitro. METHODS: A small interference RNA (siRNA) targeting to hTERT mRNA (pU6-hTERT-siRNA) was constructed. The siRNA was transfected into LoVo colon cancer cells in vivo and in vitro with Lipofectamine^TM2000. The groups of non-specific siRNA (pU6-hTERT) and non-treatment were designed as negative control and blank control,respectively. The cell growth in vitro was detected by MTT method. The effect of pU6-hTERT-siRNA on xenografts in nude mice was observed by determining the tumor size. The mRNA expression of hTERT in vitro and in vivo was detected by FQ-PCR quantitatively. The protein level of hTERT was determined by Western blotting. RESULTS: The inhibition rate of cell growth in vitro 72 h after transfection with recombinant plasmids containing hTERT-target sequences was 42.1%, significantly higher than that in control group (3.2%, P<0.01). The size of xenografts in pU6-hTERT-siRNA group was (85.9±18.7)mm³, significantly smaller than that in control group and blank group , P<0.01. The mRNA expression and the protein level of hTERT were both specifically inhibited by pU6-hTERT-siRNAs in LoVo colon cancer cells and xenografts (P<0.01). No difference between control group and blank group was observed (P>0.05).CONCLUSION: hTERT expression in LoVo colon cancer cells is inhibited significantly in vivo and in vitro by using plasmid-based siRNA. Down-regulation of hTERT expression distinctly inhibits the growth of LoVo colon cancer cells in vitro or subcutaneously transplanted in athymic mice.     

Effect of specific hTERT-siRNA on growth of human tongue cancer cells in vivo and in vitro

AIM: To investigate the effect and mechanisms of siRNA-hTERT-induced inhibition of Tca8113 tongue cancer cells in vitro and in vivo. METHODS: A small interference RNA (siRNA) targeting to hTERT mRNA (siRNA-hTERT1) was constructed. The siRNA was transfected into Tca8113 tongue cancer cells in vivo and in vitro with cationic liposome. A non-specific siRNA (siRNA-hTERT2) and non-treatment were used as negative control group and blank group. The cell growth in vitro was detected by MTT method. The cell apoptosis in vitro was analyzed by flow cytometry. The effect of siRNA-hTERT1 on xenografts in nude mice was observed by determining the tumor size. The cell apoptosis in xenografts was analyzed by Hoechst staining. The expressions of hTERT mRNA in vitro and in vivo were detected by RT- PCR. RESULTS: The inhibition rates of cell growth in vitro 72 h after siRNA-hTERT1 treatment was 47.2％, significantly higher than that in siRNA-hTERT2 treatment group (2.6％, P<0.01). The cell apoptosis rate was 27.30%±0.18% in vitro, significantly increased at 48 h after transfection of siRNA-hTERT1, compared to negative control group and blank group (P<0.01). The size of xenografts in siRNA-hTERT1 treatment group was (298.8±138.7)mm3, significantly smaller than that in siRNA-hTERT2 treatment group and blank group (495.1±151.6)mm3 and (506.8±207.4)mm3, the inhibition rate was 40.0% (P<0.01). The numbers of apoptotic cells in xenografts significantly increased after transfection of siRNA-hTERT1, compared to negative control group and blank group (P<0.01). Compared to negative control group and blank group, the expression of hTERT mRNA in Tca8113 tongue cancer cells in vitro and in vivo was inhibited by siRNA-hTERT1. CONCLUSION: siRNA-hTERT1 powerfully inhibits the growth of Tca8113 tongue cancer cells in vitro and in vivo. The specific inhibition of hTERT mRNA expression and cell apoptosis may be its main mechanisms.     

Inhibitory effect of liposomes survivin antisense oligonucleotide on human hepatic carcinoma transplanted subcutaneously in nude mice

AIM: To explore the effects of liposomes survivin antisense oligonucleotides (ASODN) on growth of human hepatic carcinoma transplanted subcutaneously in nude mice. METHODS: Nude mouse model of human hepatic cancer was established by transplantation of hepatic cancer cell line SMMC-7721/ADM subcutaneously. Models were divided randomly into six groups: control group, liposome group, sense oligonucleotide (SODN) group, 200 μg/L, 400 μg/L and 600 μg/L ASODN groups. Different treatments were given respectively. Weight and volume of subcutaneous tumors were measured, and tumor growth inhibitory rate was calculated. Morphological changes of transplanted tumor cells were observed under light microscope. The expression of Survivin was detected by immunohistochemistry. RESULTS: The growth of tumors was significantly inhibits in all ASODN groups compared with control, liposome and SODN groups (P<0.05). Volume of subcutaneous tumors decreased in a time-dependent and dosage-dependent manner (P<0.5). CONCLUSION: Survivin ASODN inhibits the growth of human hepatic carcinoma in nude mice.     

Effects of baicalin on CA46 cell xenografts in nude mice

AIM: To study the effects of baicalin on CA46 cell xenografts in nude mice. METHODS: The nude mice with CA46 cell xenografts were treated with drugs via intraperitoneal injection daily, and were divided into 5 groups: negative control group, 15 mg/kg baicalin group, 30 mg/kg baicalin group, 60 mg/kg baicalin group and 4 mg/kg etoposide (VP-16) positive control group. After 12-day treatment, the weight of CA46 cell xenografts stripped from some nude mice in the 5 groups was used to evaluate the effect of baicalin on xenograft growth in the nude mice. The apoptosis, necrosis and pathological changes of the xenograft cells were examined under light microscope and transmission electronic microscope respectively. The expression levels of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway-related proteins extracted from xenografts were determined by Western blotting. The other nude mice with CA46 cell xenografts in the 5 groups continued to be treated with the drugs until death in order to evaluate the effect of balcalin on survival time of the nude mice with CA46 cell xenografts. RESULTS: Baicalin remarkably inhibited the growth of CA46 cell xenografts, induced apoptosis and necrosis of xenograft cells, and reduced the protein expression of phospho-Akt (p-Akt), nuclear factor-kappa B (NF-κB), mammalian target of rapamycin (mTOR) and phospho-mTOR (p-mTOR) in the xenografts after 12-day treatment. Furthermore, baicalin prolonged the survival time of the nude mice with CA46 cell xenografts in a dose-dependent manner. CONCLUSION: Baicalin inhibits the growth and induces apoptosis of CA46 cell xenografts in the nude mice, and prolongs the survival time of the nude mice with CA46 cell xenografts through the mechanism of down-regulating PI3K/Akt/NF-κB and PI3K/Akt/ mTOR signaling pathways.     

Inhibitory effect of survivin antisense oligodeoxyribonucleotide combined with DDP in nude mice bearing human osteosarcoma xenograft

AIM:To investigate the feasibility and its mechanisms of improving therapeutic effect by antisense gene therapy combined with chemotherapy in osteosarcoma. METHODS:The human osteosarcoma implanted tumor model in the nude mice was established. By intratumoral injection and abdominal cavity administration, the tumor bearing mice were treated with survivin ASODN in combination with diamminedichloroplatinum (DDP) for a week. Comparison with each single-agent therapy and control group was performed in aspects such as tumor growth condition, pathological changes of tumor tissues;survivin protein expression in tumor tissues by immunohistochemistry, survivin mRNA expression levels by RT-PCR method and tumor apoptosis by Tdt-mediated dUTP nick end labeling (TUNEL). RESULTS:All nude mice survived the therapy. As compared with the control group, the antisense gene therapy group presented synchronous decrease in survivin mRNA and protein expression;all therapy group displayed tumor growth inhibition and cell apoptosis with different extent;while in contrast to single-agent therapy group, the combined therapy group showed stronger inhibition of tumor growth and abundant tumor cell apoptosis with the highest apoptotic rate. CONCLUSION:Synergistic effect was achieved by combination of DDP with ASODN that may overcome drug resistant of DDP and the combined strategy may shed new light on the cancer therapy.     

Knockdown of survivin expression using siRNA inhibits the growth of PC-3M cells

AIM: To study the inhibitory effects of survivin siRNAs on the growth of PC-3M cells. METHODS: Two pairs of DNA template coding siRNA against survivin were synthesized to construct two recombinant plasmids, pSi-sur1 and pSi-sur2. The two recombinants and the two controls, lipofectin and vacant plasmid were transfected into PC-3M cells. The expressions of survivin mRNA and protein were detected respectively by RT-PCR and Western blotting. Proliferation abilities were measured by MTT, and the cell cycle and apoptosis were assayed by FCM. RESULTS: After 72 h of transfection, the level of cell survivin mRNA in the two siRNA groups was 48%±6% (n=3) and 30%±5% (n=3) of that in lipofectin group, and expression of survivin protein were 38%±4% (n=3) and 36%±4% (n=3) respectively of that in lipofectin control. The proliferation rate of cells in pSi-sur1 and pSi-sur2 groups was also inhibited according to MTT, about 44.20%±2.08% (n=3) and 39.20%±1.93% (n=3) of that in lipofectin group. Cell numbers of G1 phase in two siRNA groups were significantly higher than that in two controls, while cells of G2 phase and S phase were much lower. Cell apoptosis was found in both siRNA groups. CONCLUSION: The two survivin siRNA significantly inhibit the expression of survivin in mRNA and protein levels, arrest the cell cycle in G1 phase, and suppress the growth of PC-3M cells and induce apoptosis in vitro.     

Apoptosis of implanted tumor of human primary gastric cancer cells in nude mice induced by resveratrol

AIM: To investigate the molecular mechanism of the apoptosis of implanted tumor of human primary gastric cancer cells in nude mice induced by resveratrol. METHODS: Human primary gastric cancer cells were planted into nude mice to establish the cancer model. Resveratrol at different doses were injected near the carcinoma on the nude mice. After treatment, transmission electron microscope and TUNEL staining method were used to detect the apoptosis of implanted tumor cells. Immunohistochemical staining and RT-PCR were used to detect the expression of apoptosis-related genes bcl-2 and bax in implanted tumor. RESULTS: Resveratrol significantly inhibited carcinoma growth when it was injected near the carcinoma. The apoptotic cells in implanted tumor induced by resveratrol were detected by transmission electron microscope and TUNEL staining, immunohistochemical staining and RT-PCR showed resveratrol inhibited bcl-2 expression and increased bax expression in human primary gastric cancer cells. CONCLUSION: Resveratrol inhibits implanted tumor of human primary gastric cancer cells in nude mice through inducing apoptosis. This apoptosis may be mediated by down-regulation of bcl-2 expression and up-regulation of bax expression.     

Construction of siRNA-survivin and GRIM-19 coexpression plasmid and its growth inhibitory effect on prostatic cancer DU145 cells

AIM: To construct the recombinant plasmid that expresses siRNA-survivin and GRIM-19 simultaneously, and to identify the validity of the recombinant plasmid and observe its effect on expression of survivin and GRIM-19 and proliferation ability of prostate cancer DU145 cells. METHODS: The recombinant plasmid coexpressing siRNA-survivin and GRIM-19 was constructed using gene cloning technique. The prostatic cancer DU145 cells were transfected with the coexpression plasmid and control plasmids. Survivin and GRIM-19 mRNA expression was detected by semi-quantitative RT-PCR. The proliferation ability affected by coexpression plasmid was measured by MTT assay. RESULTS: The coexpression plasmid pGRIM-19-si-survivin was successfully constructed according to DNA recombinant technique and identified through restriction enzyme digestion and plasmid sequencing. Compared with the mock, survivin mRNA expression levels were 0.55?0.05,0.62?0.08 and 0.35?0.05 in psi-survivin, pGRIM-19 and pGRIM-19-si-survivin groups, respectively. Compared with psi-survivin and pGRIM-19, pGRIM-19-si-survivin inhibited survivin mRNA expression markedly (P<0.05), while the expression levels of GRIM-19 mRNA were 1.93?0.14, 2.57?0.20 and 4.12?0.21 in psi-survivin, pGRIM-19 and pGRIM-19-si-survivin groups, respectively (P<0.01). Compared with pGRIM-19 group, pGRIM-19-si-survivin enhanced GRIM-19 mRNA expression more obviously (P<0.05). After transfection for 48 h, the proliferation rates were 58.0%?7.2%, 62.1%?6.1% and 50.2%?4.8% in the 3 experiment groups compared with the mock (P<0.05). After transfection for 72 h, the proliferation rate were 43.4%?4.3%, 51.3%?6.7% and 26.8%?7.1% in experiment groups compared with the mock (P<0.05). Compared with psi-survivin and pGRIM-19, pGRIM-19-si-survivin significantly inhibited the cell growth (P<0.05). CONCLUSION: Transfection of coexpression plasmid pGRIM-19-si-survivin dramatically changes the mRNA expression of survivin and GRIM-19 and inhibits the cell growth.     

Influence of hypoxia-inducible factor-1 alpha on lung cancer cell A549 growth in vitro and in vivo

AIM: To study the role of hypoxia-inducible factor-1alpha(HIF-1α) on lung cancer cells A549 growth in vitro and in vivo. METHODS: To observe the growth rate of A549 cells after HIF-1α transfected, A549 cells (1×10⁶/mouse) were inoculated subcutaneously into 20 nude mice, which were randomly divided into two groups: the control group (group A, n=10), the HIF-1α transfected group(group B, n=10). The weights of subcutaneous tumor were detected. The resected specimens were made into paraffin-embedded sections. The proliferating cell nuclear antigen (PCNA) was identified by immunohistochemistry(ISH). The expressions of HIF-1α、 apoptosis-related protein survivin and bcl-2 were analyzed by Western blot. RESULTS: The growth rates of the HIF-1α transfected lung cancer cells A549 were significantly increased, and more importantly, the HIF-1α transfected lung cancer cells A549 was able to enhance lung cancer growth in nude mice(P＜0.05). The PCNA were increased significantly in group B, compared with group A. The expressions of HIF-1α, survivin and bcl-2 in group B were increased significantly than that of group A. CONCLUSIONS: HIF-1α increases lung cancer cells A549 growth in vitro and in vivo and its mechanism may be due to promotion of proliferation and inhibition of apoptosis.     

Treatment of hepatocellular carcinoma in vitro and in vivo with yCD/TK double suicide gene driven by AFP promoter

AIM: To study the effect and mechanism of yeast cytosine deaminase/ thymidine kinase (yCD/TK) double suicide gene driven by alpha fetoprotein (AFP) promoter on hepatocellular carcinoma (HCC) in vitro and in vivo. METHODS: The expression plasmid with yCD/TK double suicide gene, which was driven by AFP promoter, was constructed. HepG2 (AFP positive) and SMMC7721 (AFP negative) human HCC cell lines were both transfected with the above-mentioned expression plasmid through cationic liposome. The cells were treated with 5-fluorocytosine (5-FC) and/or ganciclovir (GCV) at different concentrations. The cell proliferation and cell cycle phase were evaluated by MTT test and flow cytometry respectively. The effect of double suicide gene on HCC xenografts in nude mice was observed through measuring the tumor size and the number of apoptosis cells. RESULTS: The double suicide gene was expressed selectively on HepG2 cells, rather than on SMMC7721 cells. The 5-FC and/or GCV inhibited effectively the proliferation of HepG2 cells in a dose-dependent manner, but had no influence on SMMC7721 cells. The inhibitory effect on HepG2 cells among different treatments was GCV+5-FC>5-FC>GCV. In vivo, the treatments inhibited markedly the growth of HepG2 cell xenografts in nude mice, transfected with yCD/TK gene. More apoptotic cells were found in HepG2 xenografts after the treatment. However, the growth of SMMC7721 cell xenografts could not be inhibited by this double suicide gene therapy, and few apoptotic cells were found. CONCLUSION: yCD/TK double suicide gene driven by AFP promoter has a significant efficacy in treatment of AFP positive HCC. Cell apoptosis may be an important mechanism of yCD/TK double suicide gene-inhibiting the growth of HCC.     

Inhibition of survivin expression in BGC-823 cells by RNA interference

AIM: To investigate the feasibility and specificity of gastric carcinoma gene therapy by utilizing RNA interference (RNAi) to inhibit survivin expression in vitro and in vivo. METHODS: Small interference RNA (siRNA) homologous to survivin was designed. pTZU6+1-siRNA-survivin vector was constructed and transfected into BGC-823 cells. The transplanted BGC-823 tumor in nude mice was established to induce RNAi. The changes of survivin gene expression, tumor cell cycle and cell apoptosis were detected by flow cytometry, RT-PCR, Western blotting, immunochemistry and TUNEL. RESULTS: The expression of survivin was obviously inhibited by RNAi in vitro. The phase of cell cycle indicated the reduction of S phase, while G1/G0 phase increased. Cell apoptosis was obvious. Both the mRNA level and the protein expression of survivin decreased obviously. The tumor size reduced after treated with pTZU6+1-siRNA-survivin vector in vivo. The expression of survivin decreased in siRNA treatment group. In contrast, little change in control group in vitro and in vivo was observed. CONCLUSION: RNA interference down-regulates survivin gene expression, inhibits BGC-823 cell proliferation and induces cell apoptosis with good specificity, which may be a possible new approach for neoplasm gene therapy.     

NFATc1 promotes vascular generation of epithelial ovarian cancer transplanted tumor by regulating CXCR2, FGF-2 and PDGF-BB

AIM: To investigate the role of NFATc1 in vascular generation in the nude mice transplanted with human ovarian cancer SKOV3 cells. METHODS: NFATc1 expression was silenced by siRNA in SKOV3 cells. Human ovarian cancer transplantation nude mouse model was established by transplanting with SKOV3 cells in which the NFATc1 gene was silenced by siRNA technique. The expression of NFATc1, CXCR2, FGF-2 and PDGF-BB at mRNA and protein levels was determined by RT-PCR, Western blotting and immunohistochemical staining. The tumor growth, angiogenesis and lymphangiogenesis were also observed. RESULTS: Over-expression of NFATc1 was observed in human ovarian cancer tissues. The silencing of NFATc1 expression by siRNA decreased tumorigenesis of transplanted ovarian cancer cells in the nude mice, reduced tumor vascular generation and inhibited the expression of CXCR2, FGF-2 and PDGF-BB at mRNA and protein levels. CONCLUSION: NFATc1 is overexpressed in ovarian cancer. NFATc1 silencing regulates the tumor vascular generation. NFATc1 thus has potential as a therapeutic target and for use in the diagnosis and evaluating prognosis of epithelial ovarian cancer.     

iASPP on apoptosis in breast cancer cells which expressed wild type p53

AIM: To investigate the RNAi effect of the inhibitory member of the ASPP family (iASPP) on the apoptosis of human breast cancer cell MCF-7 which expressed the wild type p53 gene. METHODS: The recombinant plasmid pAd-iASPP-RNAi was transfected into MCF-7 cells. The expression of iASPP mRNA and protein was analyzed by RT-PCR and Western blotting, respectively. The cell apoptosis was detected by FCM, and then the MCF-7 cells were transplanted into nude mice to set up transplantation model. The expression of iASPP RNA and protein in transplanted neoplasm were determined by RT-PCR and Western blotting, the apoptosis index was detected by FCM at the same time. RESULTS: The results showed that the expression of iASPP descended in MCF-7 cells (mRNA 95.4% and protein 96.8%, respectively, P<0.01) and the apoptosis rate and necrosis rate of MCF-7 cells increased (P<0.01) after transfection. As treated with pAd-iASPP-RNAi, the expression of iASPP in transplantation tumor cells descended 87.4% (mRNA) and 89.2% (protein), respectively (P<0.01), and the apoptosis rate and necrosis rate increased accordingly (P<0.01, P<0.05). CONCLUSION: The inhibition of iASPP may resume the ability of p53 to induce apoptosis in breast cancer cells which is able to express wild type p53.     

Chutan-Jiedu decoction reverses resistance of lung cancer to gefitinib by processing EMT

AIM: To investigate the mechanism of Chutan-Jiedu decoction (CJD) reversing the resistance of lung cancer to gefitinib via epithelial-mesenchymal transition (EMT) pathway.METHODS: BALB/c nude mice (n=60) were selected to establish lung cancer xenograft model with human lung adenocarcinoma drug-resistant cell line H1975, which were randomly divided into 6 groups (10 mice per group):model group, gefitinib (0.04 g/kg) group, low-dose (13.52 g/kg) CJD group, middle-dose (27.04 g/kg) CJD group, high-dose (54.08 g/kg) CJD group, and combined medication group (27.04 g/kg CJD+0.04 g/kg gefitinib). The mice in each group were treated for 2 weeks before the tumor size and tumor weight were detected for the calculation of the tumor inhibitory rate. The mRNA and protein expression levels of E-cadherin, Snail and vimentin were determined by immunohistochemistry, Western blot and real-time PCR.RESULTS: Compared with model group and gefitinib group, the tumor size and the tumor weight in middle-dose CJD group, high-dose CJD group and combined medication group were decreased significantly (P<0.05). The results of immunohistochemistry, Western blot and real-time PCR showed that the expression of E-cadherin at mRNA and protein levels was increased significantly, while the expression of Snail and vimentin at mRNA and protein levels was decreased significantly (P<0.05).CONCLUSION: The growth of lung adenocarcinoma H1975 xenografts in nude mice is inhibited by CJD. In addition, the resistance of lung cancer to gefitinib is reversed. The mechanism may be related to the regulation of EMT-related protein expression.