Objective： The aim of this study was to construct THY1 eukaryotic expression plasmid ,and study its effects on ovarian cancer SKOV3 cells. Methods： The gene fragment coding for THY1 was obtained from human normal ov...Objective： The aim of this study was to construct THY1 eukaryotic expression plasmid ,and study its effects on ovarian cancer SKOV3 cells. Methods： The gene fragment coding for THY1 was obtained from human normal ovarian tissue using RT-PCR, and inserted into the eukaryotic expression plasmid pcDNA3.1 （＋） to construct the recombinant plas- mid pcDNA3.1（＋）-THY1, which was transfected into SKOV3 cells. The experimental cells were classified into three groups： SKOV3-THY1, SKOV3-Null and SKOV3. The expression of gene was measured using RT-PCR and Western blotting. The percentage of apoptotic cells and cell cycle analysis and cell proliferation were assessed by flow cytometry and MTT assay. Both SKOV3-THY1 and SKOV3-null cells were inoculated subcutaneously into nude mice to determine in vivo tumorigenicity. Results： The gene fragment of THY1 was correctly inserted into the eukaryotic expression plasmid pcDNA3.1 （＋） and veri- fied by PCR, restriction endonucleases digestion and DNA sequencing and the plasmid of pcDNA3.1（＋）-THY1 （THY1 gene overexpression） has been stably transfected into SKOV3 cells. The analysis of flow cytometry indicated that the pcDNA3.1（＋）- THY1 transfected ceils in G1 phase were significantly elevated, but in S phase were decreased. The growth of transfected cells was suppressed, and more apoptosis cells were identified in pcDNA3.1（＋）-THY1 transfectants compared with vector vehicle transfectants. The tumor suppressing activity of THY1 in SKOV3 cells was associated with inhibition of SKOV3 cellular proliferation, in vivo tumorigenesis in nude mice. Conclusion： THY1 transfection can inhibit the growth of SKOV-3 cells in vitro and in vivo. THY1 gene may play an important role in generation and development of ovarian cancers.展开更多
OBJECTIVE Epidermal growth factor receptor (EGFR) isdysregulated in many human malignancies and is a potentialtarget for therapeutic intervention, but there is a majordisagreement among researchers about both the freq...OBJECTIVE Epidermal growth factor receptor (EGFR) isdysregulated in many human malignancies and is a potentialtarget for therapeutic intervention, but there is a majordisagreement among researchers about both the frequency andpossible clinical importance of EGFR overexpression in ovariancancer. We investigated the expression and significance of theEGFR mRNA and vascular endothelial growth factor (VEGF)mRNA in ovarian carcinoma.METHODS Reverse transcription polymerase chain reaction(RT-PCR) was employed to determine the expression of EGFRmRNA and VEGF mRNA in 79 ovarian specimen (including15 normal, 13 benign and 51 malignant, from 79 patients). Therelationship between EGFR and VEGF expression was analyzed.RESULTS The positive rates of the expression of EGFR mRNAand VEGF mRNA were significantly higher in the patientswith ovarian carcinoma than those in both the patients withbenign ovarian tumors and in the normal controls. There wascorrelation between EGFR mRNA expression and clinical stages.The positive rate of the expression of EGFR mRNA in Stage Ⅲ-Ⅳ was higher than that in Stage Ⅰ-Ⅱ of ovarian carcinoma (P <0.05). The expression of VEGF mRNA was correlated with theclinical stages and lymph node metastasis. The expression levelsof VEGF mRNA in Stage Ⅲ-Ⅳ and in the group with lymph nodemetastasis were significantly higher than those in Stage Ⅰ-Ⅱ and inthe group without lymph node metastasis, respectively (P < 0.05).The expression of EGFR mRNA was positively correlated with theexpression of VEGF mRNA (r = 0.438, P < 0.05).CONCLUSION The expressions of EGFR mRNA and VEGFmRNA are positively correlated to the occurrence of ovariancarcinoma and its metastasis. The detection of EGFR and VEGFmay be helpful for the targeted chemotherapy.展开更多
基金Supported by a grant from the Medical Research Foundation of Guang-dong Province No A2008099
文摘Objective： The aim of this study was to construct THY1 eukaryotic expression plasmid ,and study its effects on ovarian cancer SKOV3 cells. Methods： The gene fragment coding for THY1 was obtained from human normal ovarian tissue using RT-PCR, and inserted into the eukaryotic expression plasmid pcDNA3.1 （＋） to construct the recombinant plas- mid pcDNA3.1（＋）-THY1, which was transfected into SKOV3 cells. The experimental cells were classified into three groups： SKOV3-THY1, SKOV3-Null and SKOV3. The expression of gene was measured using RT-PCR and Western blotting. The percentage of apoptotic cells and cell cycle analysis and cell proliferation were assessed by flow cytometry and MTT assay. Both SKOV3-THY1 and SKOV3-null cells were inoculated subcutaneously into nude mice to determine in vivo tumorigenicity. Results： The gene fragment of THY1 was correctly inserted into the eukaryotic expression plasmid pcDNA3.1 （＋） and veri- fied by PCR, restriction endonucleases digestion and DNA sequencing and the plasmid of pcDNA3.1（＋）-THY1 （THY1 gene overexpression） has been stably transfected into SKOV3 cells. The analysis of flow cytometry indicated that the pcDNA3.1（＋）- THY1 transfected ceils in G1 phase were significantly elevated, but in S phase were decreased. The growth of transfected cells was suppressed, and more apoptosis cells were identified in pcDNA3.1（＋）-THY1 transfectants compared with vector vehicle transfectants. The tumor suppressing activity of THY1 in SKOV3 cells was associated with inhibition of SKOV3 cellular proliferation, in vivo tumorigenesis in nude mice. Conclusion： THY1 transfection can inhibit the growth of SKOV-3 cells in vitro and in vivo. THY1 gene may play an important role in generation and development of ovarian cancers.
文摘OBJECTIVE Epidermal growth factor receptor (EGFR) isdysregulated in many human malignancies and is a potentialtarget for therapeutic intervention, but there is a majordisagreement among researchers about both the frequency andpossible clinical importance of EGFR overexpression in ovariancancer. We investigated the expression and significance of theEGFR mRNA and vascular endothelial growth factor (VEGF)mRNA in ovarian carcinoma.METHODS Reverse transcription polymerase chain reaction(RT-PCR) was employed to determine the expression of EGFRmRNA and VEGF mRNA in 79 ovarian specimen (including15 normal, 13 benign and 51 malignant, from 79 patients). Therelationship between EGFR and VEGF expression was analyzed.RESULTS The positive rates of the expression of EGFR mRNAand VEGF mRNA were significantly higher in the patientswith ovarian carcinoma than those in both the patients withbenign ovarian tumors and in the normal controls. There wascorrelation between EGFR mRNA expression and clinical stages.The positive rate of the expression of EGFR mRNA in Stage Ⅲ-Ⅳ was higher than that in Stage Ⅰ-Ⅱ of ovarian carcinoma (P <0.05). The expression of VEGF mRNA was correlated with theclinical stages and lymph node metastasis. The expression levelsof VEGF mRNA in Stage Ⅲ-Ⅳ and in the group with lymph nodemetastasis were significantly higher than those in Stage Ⅰ-Ⅱ and inthe group without lymph node metastasis, respectively (P < 0.05).The expression of EGFR mRNA was positively correlated with theexpression of VEGF mRNA (r = 0.438, P < 0.05).CONCLUSION The expressions of EGFR mRNA and VEGFmRNA are positively correlated to the occurrence of ovariancarcinoma and its metastasis. The detection of EGFR and VEGFmay be helpful for the targeted chemotherapy.
