共查询到20条相似文献，搜索用时 31 毫秒
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AIM: To investigate the effects of p27kip1 gene on DNA replication and protein synthesis in esophageal carcinoma cells. METHODS: Recombinant adenovirus Ad-p27kip1 was constructed and transfected into esophageal carcinoma cell EC-9706, and its effects on DNA replication, protein synthesis and the growth of esophageal carcinoma cells were explored by means of cell growth count, [3H]-TdR, [3H]-Leucine incorporation method and flow cytometry. RESULTS: The growth of esophageal carcinoma cells was inhibited obviously. The radioactive intensity of -TdR and -Leucine in Ad-p27kip1 group decreased significantly than that in control group after EC-9706 transfection (P<0.01), while the multiplication of esophageal carcinoma cell was inhibited obviously. CONCLUSION: Ad-p27kip1 inhibited the multiplication of the esophageal carcinoma cells, which may be related to its suppressive function for DNA replication and protein synthesis. 相似文献
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AIM: To investigate expression of CD44s in lung cancer and it's clinical significance. METHODS: A total of 117 primary lung cancer from patients were examined for CD44s expression by immunohistochemical staining. RESULTS: CD44s mostly expressed in non-small cell lung cancer (NSCLC) but not in small ecll lung cancer (SCLC), and squamous cell carcinoma(SCC) showed much stronger expression of CD44s than adenocarcinoma(ADC)(P＜0.05). In comparison of the lung cancer with/ without lymph node metastasis, the latter showed stronger expression of CD44s(P＜0.01). According to TNM, there was a distinct statistic difference between early stage and advanced stage(P＜0.05). CONCLUSION: CD44s might be a better indicant in histological classification of lung cancer, lymph node metastasis, clinical stage and prognosis. 相似文献
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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. 相似文献
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The high morbidity and mortality of pulmonary carcinoma, especially in male patient, are still the unsolved problems. Recently, more and more reports showed that gene therapy could have potential application in the treatment of the disease. In this article, some progresses in the gene therapy of pulmonary carcinoma, such as tumor suppressor gene, drug sensitive gene, antisense gene, multi-drug resistance gene, immune gene as well as anti-angiogenesis gene, were discussed. 相似文献
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AIM: To explore the relationship between the expression of transforming growth factor-β1 (TGF-β1) and the pathogenesis of lung carcinoma, and the influence of radiotherapy on plasma TGF-β1 level of patients with lung carcinoma. METHODS: By immunohistochemical method, the expression of TGF-β1 was examined. An enzyme-linked immunoadsorbent assay (ELISA) was used to quantify the plasma TGF-β1 levels in different time as before radiotherapy, at the end of radiotherapy, and at the time of follow-up 6 months after radiotherapy, respectively. The changes of quantity of TGF-β1 in different time above were analysed statistically. RESULTS: There was a significant increase in TGF-β1 expression in the carcinoma compared with normal lung tissue. The mean TGF-β1 level in the 39 lung carcinoma patients before radiotherapy was (11.0±1.5) μg/L, which was significantly higher than control group (3.8±0.2 μg/L) (P＜0.05); but the plasma TGF-β1 level after radiotherapy was significantly lower (5.6±0.5 μg/L) compared with the level before radiotherapy (P＜0.05), and there was not significant differences compared with control group (P＞0.05). At the time of follow-up 6 months, the patients of lung carcinoma had a significantly higher plasma TGF-β1 level (11.3±1.2 μg/L) compared with the level at the end of radiotherapy (P＜0.05), but there was not significant differences compared with before radiotherapy (P＞0.05). Not significant difference was found in TGF-β1 levels among different histologic types. CONCLUSION: These data demonstrated that TGF-β1 was associated with the pathogenesis of lung carcinoma, and it may be a useful tumor marker in patients with lung carcinoma. 相似文献
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AIM: To investigate the change of chemosensitivity of esophageal carcinoma cells before and after induction by radiation,and to detect the excision repair cross complementation group 1 (ERCC1) gene before and after induction,and further to analyze the relationship between ERCC1 expression and chemosensitivity.METHODS: The esophageal carcinoma cell EC9706 was radiated repeatedly by a long-term,intermittent ［60Co］-γ radiation to induce the radioresistance esophageal carcinoma cell EC9706-R.The IC50 and resistant index (RI) of EC9706 and EC9706-R were detected by MTT assay.The expression of ERCC1 was examined by immunocytochemistry and RT-PCR.RESULTS: The IC50 of EC9706 and EC9706-R cells to cisplatin were (1.480±0.012) mg/L and (1.836±0.008) mg/L,respectively (P<0.05),and the RI was 1.240±0.015.The tinctorial scores of ERCC1 protein in EC9706 and EC9706-R were 2.838±0.055 and 2.898±0.039 (P>0.05),and the relatively quantities (IDV) of ERCC1 mRNA in EC9706 and EC9706-R cells were 1.168±0.068 and 1.143±0.089 (P>0.05).SP and RT-PCR did not display visible difference in protein level and mRNA level.CONCLUSION: The chemosensitivity of EC9706-R cells to cisplatin is decreased compared with EC9706 cells,but ERCC1 expression does not change with the radioresistance and chemoresistance. 相似文献
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AIM: To evaluate the changes of serum total prostate specific antigen (T-PSA), free prostate specific antigen (F-PSA) and the ratio of F-PSA to T-PSA (F/T) in patients with prostate cancer and its clinical significance. METHODS: The concentrations of T-PSA and F-PSA in serum were measured by micropartical enzyme immunoassay (MEIA) using AxSYM System, and the F/T ratio was calculated. RESULTS: Before operation, the concentrations of T-PSA and F-PSA in patients with PCa were much higher and F/T ratio was significantly lower than that in patients with benign prostate hyperplasia (BPH). T-PSA and F-PSA levels decreased, but F/T ratio increased after operation in PCa and BPH. F/T ratio in 83.5% PCa and 6.5% BPH was less than 0.16. To diagnosis PCa, the sensitivity of F/T ratio was 83.5%, and the specificity was 86.7%. CONCLUSION: Serum T-PSA, F-PSA and F/T ratio are important parameters for the early diagnosis of prostate cancer. 相似文献
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LIU Lan-ying YANG Kun ZHU Zheng-yu LIU Yu-chuan YU Xi-yong TANG Jian YIN Wei MA Jian-quan GU Jun 《园艺学报》2000,16(5):389-392
AIM: To study cellular and molecular mechanisms of cardiac development associated genes expression and its function during early stage cardiomyogenesis. METHODS: (1) Mouse embryonic stem cells (ESC) line D3 culture.(2) Inductive culture of ESC differentiated into cardiac myocytes in vitro. (3) Identification of ESC-derived cardiac myocytes: RNA isolation; synthesis of specific primer and RT-PCR; Label of RT-PCR products with dATP as probes, purifyed by sephadex G-50 columns, determined the yield of DNA. RNA dot hybridization. RESULTS: 80% of ESC differentiated into cardiomyocytes by improved conditional medium. Cardiomyocytes contracted in a synchronous manner. The results of RT-PCR and RNA blot showed that cardiac genes were expressed abundantly and specifically during the early cardiomyogenesis. CONCLUSIONS: ESC were able to be differentiate into cardiomyocytes. Different concentrations and components of RA, DMSO and FCS affected ESC cardiomyogenesis in vitro. The optimal result obtained was from the conditional medium, a mixturce of 2 nmol/L retinoic acid (RA), 0.6% dimethyl sulfoxide (DMSO) and 20% fetal calf serum (FCS). 相似文献
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AIM: To investigate the transfection efficiency of mouse liver with non-viral vector containing manganese superoxide dismutase (Mn-SOD) gene. METHODS: The eukaryotic expression vector, gWiz/Mn-SOD, encoding human manganese superoxide dismutase was constructed. The plasmids of gWiz/Mn-SOD were mixed with cationic lipids, followed by injection into mice via branch of superior mesenteric vein, to induce Mn-SOD over-expression in murine liver detected by RT-PCR, Western blotting, SOD activity and immunohistochemical staining. RESULTS: gWiz/Mn-SOD transfection resulted in the obvious expression of exogenous Mn-SOD mRNA and protein in hepatic tissues at 8 hours after injection, and elevated mitochondria SOD activity 8.4 times in transfected hepatocytes than that in non-transfected cells at 72 hours after injection. It was showed that nearly 70% of mouse hepatocytes was obviously Mn-SOD positive after transfection. CONCLUSION: High expression efficiency of Mn-SOD gene in mouse liver is achieved safely, by injection of gWiz/Mn-SOD and cationic lipid mixture into branch of superior mesenteric vein. 相似文献
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AIM: To study the effect and mechanism of chlorophyllin (CHL) inhibiting HT29 cells. METHODS: IC50 value and growth curve of HT29 cells were detected with MTT method. Apoptosis was detected with Wright-Giemsa staining, FCM and DNA electrophoresis. Telomerase was detected by PCR-ELISA, and protein and mRNA expression of COX-2 gene were detected through RT-PCR and Western blot. RESULTS: CHL inhibited the growth of HT29 in a dose-dependent manner. CHL blocked HT29 cells in G1 phase but did not induce apoptosis. Different concentration of CHL inhibits the expression of telomerase and COX-2 in HT29 cells. CONCLUSION: CHL inhibited the growth of HT29 cells by inhibiting the expression of telomerase and COX-2 and blocking cells in G1 phase. 相似文献
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AIM: To evaluate the significance of cytokeratin19 (CK19) expression in diagnosing micrometastases in lymph nodes in patients with laryngeal carcinoma. METHODS: Forty cases of laryngeal carcinoma together with 163 lymph nodes were studied by the staining with hematoxylin and eosin(HE)and immunostaining with antibody against cytokeratin 19 (CK19) on histological sections of the primary tumor and regional lymph nodes. RESULTS: In all cases, CK19 was positively expressed in the primary tumor. Among 163 lymph nodes, metastases were confirmed by HE in 23 lymph nodes, and in 42 lymph nodes staining with immunohistochemistry, micrometastases were found in 19 lymph nodes. Micrometastases in the lymphonodes were significantly related to the T staging. CONCLUSION: Immunohistochemical staining is a valuable method for the detection of node micrometastases in patients with laryngeal carcinoma. 相似文献
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ZHANG Zai-yun LI Xiao-mei WANG Juan-dong SUN Jian-hua JIANG Yu-hua PAN Xiang-lin 《园艺学报》2011,27(9):1758-1761
AIM: To study the effect of interleukin 18( IL-8 ) gene modification on anti-tumor activity induced by lung cancer cell-derived exosomes.METHODS: Exosomes isolated from the supernatants of IL-18 gene-modified NCI-H460 lung cancer cells (IL-18/H460), pcDNA3.1+ vector-modified cancer cells (DNA3.1/H460) and non-modified NCI-H460 lung cancer cells (NCI-H460) were observed under transmission electron microscope.The expression of heat-shock protein 70(HSP70),human leukocyte antigen(HLA) and IL-18 were determined by Western blotting.T lymphocytes were activated by exosomes or exosome-pulsed dendritic cells(DCs).The activity of T cells for killing lung cancer cells were detected by lactate dehydrogenase (LDH) method.The killing rates were calculated and compared.RESULTS: Exosomes showed typical morphous under transmission electron microscope.The protein levels of HSP70 and HLA were detected in the exosomes of all 3 groups, and IL-18 protein was only observed in IL-18/H460 group.The killing rates of exosome-activated T cells in IL-18/H460 group with the ratio of effector cell to target cell at 25∶ 1, 10∶ 1 and 5∶ 1 were (38.45±5.42)%, (25.17±3.94)% and (11.75±3.22)%, respectively.The killing rates of exosome-pulsed DC-activated T cells in this group were (89.05±4.06)%, (64.97±6.02)% and (40.16±4.98)%, respectively.The killing rates in IL-18/H460 group were higher than those in DNA3.1/H460 group and NCI-H460 group.The anti-tumor efficacy of exosome-pulsed DC－activated T cells was stronger than that of exosome-activated T cells.CONCLUSION: IL-18 gene modification enhances the anti-tumor activity induced by NCI-H460 lung cancer cell-derived exosomes. 相似文献
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AIM: To study the expression and its kinetics of rice phenylalanine ammonia-lyase gene encoding into E. coli as the basis of treatment for phenylketouria. METHODS: The phenylalanine ammonia-lyase-1-cDNA(rPAL-1-cDNA) from rice was recombined into E. coli high expression vector pET-28c and transformed into E. coli host strain BL21DE3. Engineering bacteria was then inducted by isopropyl-β-D-thiogalactoside (IPTG) for 1, 3, 5, 7 hours, in order to obtain high level expression. RESULTS: After induction, the expression level of fusion protein was 21.40%, 30.60%, 35.40%, 35.43% respectively. The fusion protein exhibited a band of 78.6 kD on SDS-PAGE analysis, but was not found in controls.The target protein was mainly existed in the form of inclusion body. CONCLUSION:Rice PAL gene expressing E. coli was established by gentic engineering technique. 相似文献
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WANG Xiao-hui LU Jian YU Xiao-ling CHEN Guang-chun ZHANG Jin-shan SUN Ying-hao 《园艺学报》2000,16(1):8-11
AIM and METHODS: To learn more about the mechanism of prostate cancer (PC) development and progression to androgen independence, the exons BH of the androgen receptor (AR) gene of forty-five patients with prostate cancer, six puncture tissues and thirty-nine slide tissues, were analyzed by polymerase chain reaction-single strand conformation technique (PCR-SSCP). RESULTS: Seven abnormal mobility shifts were found in five patients by PCR-SSCP. Combining the method with direct DNA cycle sequencing, two distinct missense (Glu872Gln, Met886Ile) point mutations were identified in puncture tissues from two patients of advanced prostate cancer with distant metastasis. These two point mutations represented two novel mutations. CONCLUSION: AR gene mutations might play an important role in the development and progression of prostate cancer. 相似文献
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AIM: To investigate the possible effect of different Latent membrane protein 1 (LMP1) variants on proliferation characteristics of a human well-differentiated nasopharyngeal carcinoma (NPC) cell line CNE1. METHODS: The plasmids which carried EBV-LMP1 gene cloned from B95-8 lymphocyte (B95-8-LMP1) and NPC tissues (CAO-LMP1) were introduced into CNE1 by liposomal transfection. Expression of LMP1 in CNE1 was identified by RT-PCR and Western blot, respectively. Different Effects of the two EBV-LMP1 variants on proliferation characteristics of CNE1 including growth curve, cell cycle distribution and clony-forming efficiency were investigated by means of crystal violet staining proliferation assay, flow cytometry and plastic plate clony-forming assay, respectively. RESULTS: Two transfected cell lines stably expressing different LMP1 variants were established successfully. Either of the two LMP1 variants increased CNE1 growth rate, proliferation index (PI) and clony-forming rate significantly and CAO-LMP1 had more significant growth-promoting effect on CNE1 than B95-8-LMP1. CONCLUSION: It can be concluded from the study that CAO-LMP1 promotes CNE1 proliferation more effectively than B95-8-LMP1. 相似文献
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AIM: To study the expression of hepatitis B virus core gene in Pichia pastoris and to obtain high-level expressed recombinant HBcAg with good immunoreactivity and high specificity. METHODS: HBV core gene was amplified by PCR from plasmid pHBV1 which contained HBV whole DNA sequence. The PCR product was cloned into pGEM-T vector by TA cloning strategy. After confirmed by DNA sequence analysis, the gene of interest was inserted into the yeast expression vector pPIC9. The recombinant plasmid pPIC9-cAg was constructed and transformed into GS115 by electroporation. The recombinant yeast GS115 was induced by 0.5% methanol. The expressed product was analysed by SDS-PAGE,Western blot and ELISA. RESULTS: The restriction analysis and DNA sequence analysis proved that HBV core gene had already been cloned to yeast expression plasmid pPIC9. The expressed HBcAg existed in SDS-PAGE. Good immunoreactivity and high specificity of the recombinant HBcAg have been proved by ELISA and Western blot. The titre of the recombinant HBcAg in the cell lysate was 1∶12 800. CONCLUSION: The recombinant plasmid pPIC9-cAg was successfully constructed. The recombinant HBcAg with good immunoreactivity and high specificity was successfully expressed in Pichia pastoris expression system and can be applied to further developing HBcAb immunoassay. 相似文献