Objective To investigate the role of extracellular regulated kinase (ERK1/2) pathway in cisplatin-induced apoptosis in human ovarian carcinoma cells. Methods Cisplatin-induced apoptosis were stained with DAPI and was ...Objective To investigate the role of extracellular regulated kinase (ERK1/2) pathway in cisplatin-induced apoptosis in human ovarian carcinoma cells. Methods Cisplatin-induced apoptosis were stained with DAPI and was assessed microscopically in human epithelial adenocarcinoma ovarian cell line SKOV3 cells. ERK activation was determined by Western blotting using an anti-phospho-ERK antibody to detect ERK activity. The effect of PD98059 on ERK activity induced by cisplatin was detected by MTT assay. Results Marked apoptosis of SKOV3 cells resulted from 48 hours treatment with 20 μg/mL cisplatin. Strong activation of ERK was led to by 15 μg/mL cisplatin. Dose response and time course of cisplatin induced apoptosis in SKOV3 cells. Cisplatin-induced ERK activation occurred at 12 hours and increased to highest induction at 24 hours by Western blotting. The effect of PD 98059 on ERK activity induced by cisplatin at the concentration of 100 μmol/L PD 98059. Statistically significant decreased in cell survival were observed with 100 μmol/L PD 98059 at 15 and 20 μg/mL cisplatin (P< 0.05). Conclusions Cisplatin activates the ERK signaling pathway in ovarian cancer cell line SKOV3. Inhibition of ERK acti-vity enhances sensitivity to cisplatin cytotoxity in ovarian cancer cell line SKOV3. Evaluation of ERK activity could be useful in predicting which ovarian cancer will response most favorably to cisplatin therapy.展开更多
Objective: To investigate the feasibility of obtaining mature spermatozoa for intracytoplasmic sperm injection (ICSI) by testicular fine needle aspiration (TEFNA) in men diagnosed non-obstructive azoospermia. Methods:...Objective: To investigate the feasibility of obtaining mature spermatozoa for intracytoplasmic sperm injection (ICSI) by testicular fine needle aspiration (TEFNA) in men diagnosed non-obstructive azoospermia. Methods: TEFNA was performed in 121 patients with a mean of 15 punctures and aspirations from each testis with a #23 butterfly needle connected to a 20 mL syringe with an aspiration handle. Results: One hundred and twenty-one patients underwent 176 TEFNA cycles. Testicular sperm were recovered in 56.3 % (99/176) cycles from 57 % (69/121) of patients. The sperm recovery rate was 46.7 % (21/45) in patients with Sertoli cell-only syndrome, 45.7 % (16/35) in patients with maturation arrest, 96.1 % (25/26) in patients with hypospermatogenesis and 63.6 % (7/11) in patients of non-mosaic Klinefelter's syndrome as judged by testicular histology. No sperm were found in 3 cases with post-irradiation fibrosis and one, after resection and chemotherapy of unilateral testicular cancer. In 87 cycles of ICSI using the husbands' sperm, 591 mature oocytes were injected, 218 (36.9 %) were normally fertilized and 202 embryos developed; 178 were transferred in 62 cycles resulting in 26 pregnancies (41.9 %) with 44 gestational sacs (implantation rate: 24.7 %). Conclusion: TEFNA was an efficient, easy to learn, safe and well tolerated treatment in patients with non-obstructive azoospermia.展开更多
Objective:To evaluate the inhibitory effect of somatostatin analogue RC-160 on the growth of human endometrial cancer cells (HEC-1A) in vitro.Method:RT-PCR was used to examine the existence of somatostatin receptors o...Objective:To evaluate the inhibitory effect of somatostatin analogue RC-160 on the growth of human endometrial cancer cells (HEC-1A) in vitro.Method:RT-PCR was used to examine the existence of somatostatin receptors on the HEC-1A cells. The anti-proliferative effect of RC-160 on the growth of HEC-1A cells was detected by using BrdU incorporation test.TUNEL staining was used to find out whether apoptosis was involved in the inhibitory process.Result:All the five somatostatin receptor subtypes were demonstrated in HEC-1A cells.RC-160 reduced the HEC-1A cell growth stimulated by serum in a dose-dependent manner.The effect was maximal at the concentration of 10 -5 M after 48 hours’ treatment.No apoptosis was detected.Conclusion:Somatostatin analogue RC-160 can inhibit the proliferation of endometrial carcinoma cell line HEC-1A through binding to the somatostatin receptors on the cells.It seems that apoptosis is not mainly responsible for the inhibition.展开更多
OBJECTIVE To evaluate the efficiency of surgery plus radiotherapy and chemotherapy versus radiotherapy plus chemotherapy in the treatment of older patients with stage I or II cervical carcinoma and to seek suitable tr...OBJECTIVE To evaluate the efficiency of surgery plus radiotherapy and chemotherapy versus radiotherapy plus chemotherapy in the treatment of older patients with stage I or II cervical carcinoma and to seek suitable treatment for such patients. METHODS The clinical data of 179 elderly women with stage la or lib cervical cancer were analyzed retrospectively. One hundred and thirty-four cases underwent radical hysterectomy followed by adjuvant radiotherapy and/or chemotherapy (Group 1). Forty-five cases underwent radiation therapy plus adjuvant chemotherapy (Group 2). RESULTS The 5-year survival rates in group 1 and group 2 were 78.3% and 49.1%(P=0.04), respectively. The incidence of complications in group1 was 47.0%. Three patients died of complications after radical hysterectomy. The incidence of complications in group 2 was 75.6%. CONCLUSION Elderly patients with stage I or II cervical carcinoma should receive an operation if possible. In addition they should receive adjuvant treatments according to their personal conditions, and be treated with appropriate adjuvant chemo-and/or radiotherapy.展开更多
Objective.The aim of the study was to evaluate the prog-nostic significance of tumor geogra phy,defined as exo-phytic or barrel -shaped growth,in b ulky(>4cm)cer-vical cancer.Methods.Four hundred women with cervica...Objective.The aim of the study was to evaluate the prog-nostic significance of tumor geogra phy,defined as exo-phytic or barrel -shaped growth,in b ulky(>4cm)cer-vical cancer.Methods.Four hundred women with cervical cancer,treated by primary radical h ysterectomy between January 1984and November 2000,were followed in a prospective cohort study.Clinical and pathology data were stored in a databank and the clinical protocolwas un-changed during the study except for t he amendment of ad-ditional indications of postoperative radiation in 1997.The assessment of tumor geography was based on pelvic exam-ination at the time of tumor staging o r radical hysterectomy or from the pathology report.Surviv al probabilities were calculated by the Kaplan -Meier meth od and compared with the log -rank test.Results.The mean age of the patients was 45years and the mean follow -up duration 48months.Tumors were of squamous cell type in 291patients(73%).Lymph node metastases were present in 91pa-tients(24%)and postoperative radiation was giv en in 179patients(45%).In 291patients,tumor diameter was<4cm;in 58patients,the tumor was defi ned as bulky exo-phytic and in 51patients as bulky barrel shaped.There were no differences among these thre e groups in terms of operating time,blood loss during su rgery or complications at3or 6months postoperatively.Bulky exophytic tumors had an identical overall survival as com pared to small -diam-eter(<4cm)tumors.The overall survival(OS)of bulky barrelshaped tumors was significan tly worse(P<10 -4 ).The same was found for disease -free s urvival(DFS).Conclusion.Bulky exophytic cervic al cancer has an iden-tical surgical morbidity,overall a nd disease -free survival as compared to nonbulky(<4cm)cervical cancer.In view of these identical characteris tics,primary surgical treatment should be considered for p atients with bulky ex-ophytic cervical cancer.展开更多
The second mitochondria-derived activator of caspases (Smac) is a novelproapoptotic gene, which plays an important role in the apoptosis-inducing effects of irradiation ontumor cells. The purpose of this study was to ...The second mitochondria-derived activator of caspases (Smac) is a novelproapoptotic gene, which plays an important role in the apoptosis-inducing effects of irradiation ontumor cells. The purpose of this study was to investigate the effects of extrinsic Smac genetransfer and its over-expression in radiotherapeutic sensitivities of cervical cancer cells. Afterthe Smac gene was transferred into the cervical cancer cell line HeLa, subcloned cells were obtainedby persistent G418 selection. Cellular Smac gene expression was detected by RT-PCR and Westernblot, while in vitro cell viabilities were detected by trypan blue staining assay. After treatmentwith X-ray irradiation, cellular radiotherapeutic sensitivities were investigated by tetrazoliumbromide colorimetry. Cellular apoptosis and its rate were determined by electronic microscopy,annexin V-FITC and propidium iodide staining flow cytometry. The expression and activities ofcellular caspase-3 were assayed by Western blot and colorimetry. Smac mRNA and protein levels inHeLa/Smac cells and the selected subclone cell line of cervical cancer were significantly higherthan those of HeLa (P <0. 01). There was no significant difference in cellular viabilities betweenthem (P > 0. 05 ) . However, after irradiation with 8 Gy X-ray, growth activities of HeLa/ Smac werereduced by 22.42% (P < 0. 01). When compared with those of HeLa, partial HeLa/Smac cells presentedcharacteristic morphological changes of apoptosis under electronic microscope, with higher apoptosisrates (16. 4% vs. 6. 2% , P < 0. 01 ) ; the caspase-3 expression levels in HeLa/Smac cells wereimproved significantly (P <0. 01) , while its activities were increased by 3. 42 times (P <0. 01).Stable transfer of the extrinsic Smac gene and its over-expression in cervical cancer cell linecould significantly enhance the expression and activities of cellular caspase-3 and ameliorateapoptosis-inducing effects of irradiation on cancer cells, which was a novel strategy to improveradiotherapeutic effects on cervical cancer.展开更多
基金Supported by Heilongjiang Province Natural Science Funds (D03-55) and Heilongjiang Province Sanitary Bureau Science Funds (2003-003)
文摘Objective To investigate the role of extracellular regulated kinase (ERK1/2) pathway in cisplatin-induced apoptosis in human ovarian carcinoma cells. Methods Cisplatin-induced apoptosis were stained with DAPI and was assessed microscopically in human epithelial adenocarcinoma ovarian cell line SKOV3 cells. ERK activation was determined by Western blotting using an anti-phospho-ERK antibody to detect ERK activity. The effect of PD98059 on ERK activity induced by cisplatin was detected by MTT assay. Results Marked apoptosis of SKOV3 cells resulted from 48 hours treatment with 20 μg/mL cisplatin. Strong activation of ERK was led to by 15 μg/mL cisplatin. Dose response and time course of cisplatin induced apoptosis in SKOV3 cells. Cisplatin-induced ERK activation occurred at 12 hours and increased to highest induction at 24 hours by Western blotting. The effect of PD 98059 on ERK activity induced by cisplatin at the concentration of 100 μmol/L PD 98059. Statistically significant decreased in cell survival were observed with 100 μmol/L PD 98059 at 15 and 20 μg/mL cisplatin (P< 0.05). Conclusions Cisplatin activates the ERK signaling pathway in ovarian cancer cell line SKOV3. Inhibition of ERK acti-vity enhances sensitivity to cisplatin cytotoxity in ovarian cancer cell line SKOV3. Evaluation of ERK activity could be useful in predicting which ovarian cancer will response most favorably to cisplatin therapy.
文摘Objective: To investigate the feasibility of obtaining mature spermatozoa for intracytoplasmic sperm injection (ICSI) by testicular fine needle aspiration (TEFNA) in men diagnosed non-obstructive azoospermia. Methods: TEFNA was performed in 121 patients with a mean of 15 punctures and aspirations from each testis with a #23 butterfly needle connected to a 20 mL syringe with an aspiration handle. Results: One hundred and twenty-one patients underwent 176 TEFNA cycles. Testicular sperm were recovered in 56.3 % (99/176) cycles from 57 % (69/121) of patients. The sperm recovery rate was 46.7 % (21/45) in patients with Sertoli cell-only syndrome, 45.7 % (16/35) in patients with maturation arrest, 96.1 % (25/26) in patients with hypospermatogenesis and 63.6 % (7/11) in patients of non-mosaic Klinefelter's syndrome as judged by testicular histology. No sperm were found in 3 cases with post-irradiation fibrosis and one, after resection and chemotherapy of unilateral testicular cancer. In 87 cycles of ICSI using the husbands' sperm, 591 mature oocytes were injected, 218 (36.9 %) were normally fertilized and 202 embryos developed; 178 were transferred in 62 cycles resulting in 26 pregnancies (41.9 %) with 44 gestational sacs (implantation rate: 24.7 %). Conclusion: TEFNA was an efficient, easy to learn, safe and well tolerated treatment in patients with non-obstructive azoospermia.
文摘Objective:To evaluate the inhibitory effect of somatostatin analogue RC-160 on the growth of human endometrial cancer cells (HEC-1A) in vitro.Method:RT-PCR was used to examine the existence of somatostatin receptors on the HEC-1A cells. The anti-proliferative effect of RC-160 on the growth of HEC-1A cells was detected by using BrdU incorporation test.TUNEL staining was used to find out whether apoptosis was involved in the inhibitory process.Result:All the five somatostatin receptor subtypes were demonstrated in HEC-1A cells.RC-160 reduced the HEC-1A cell growth stimulated by serum in a dose-dependent manner.The effect was maximal at the concentration of 10 -5 M after 48 hours’ treatment.No apoptosis was detected.Conclusion:Somatostatin analogue RC-160 can inhibit the proliferation of endometrial carcinoma cell line HEC-1A through binding to the somatostatin receptors on the cells.It seems that apoptosis is not mainly responsible for the inhibition.
文摘OBJECTIVE To evaluate the efficiency of surgery plus radiotherapy and chemotherapy versus radiotherapy plus chemotherapy in the treatment of older patients with stage I or II cervical carcinoma and to seek suitable treatment for such patients. METHODS The clinical data of 179 elderly women with stage la or lib cervical cancer were analyzed retrospectively. One hundred and thirty-four cases underwent radical hysterectomy followed by adjuvant radiotherapy and/or chemotherapy (Group 1). Forty-five cases underwent radiation therapy plus adjuvant chemotherapy (Group 2). RESULTS The 5-year survival rates in group 1 and group 2 were 78.3% and 49.1%(P=0.04), respectively. The incidence of complications in group1 was 47.0%. Three patients died of complications after radical hysterectomy. The incidence of complications in group 2 was 75.6%. CONCLUSION Elderly patients with stage I or II cervical carcinoma should receive an operation if possible. In addition they should receive adjuvant treatments according to their personal conditions, and be treated with appropriate adjuvant chemo-and/or radiotherapy.
文摘Objective.The aim of the study was to evaluate the prog-nostic significance of tumor geogra phy,defined as exo-phytic or barrel -shaped growth,in b ulky(>4cm)cer-vical cancer.Methods.Four hundred women with cervical cancer,treated by primary radical h ysterectomy between January 1984and November 2000,were followed in a prospective cohort study.Clinical and pathology data were stored in a databank and the clinical protocolwas un-changed during the study except for t he amendment of ad-ditional indications of postoperative radiation in 1997.The assessment of tumor geography was based on pelvic exam-ination at the time of tumor staging o r radical hysterectomy or from the pathology report.Surviv al probabilities were calculated by the Kaplan -Meier meth od and compared with the log -rank test.Results.The mean age of the patients was 45years and the mean follow -up duration 48months.Tumors were of squamous cell type in 291patients(73%).Lymph node metastases were present in 91pa-tients(24%)and postoperative radiation was giv en in 179patients(45%).In 291patients,tumor diameter was<4cm;in 58patients,the tumor was defi ned as bulky exo-phytic and in 51patients as bulky barrel shaped.There were no differences among these thre e groups in terms of operating time,blood loss during su rgery or complications at3or 6months postoperatively.Bulky exophytic tumors had an identical overall survival as com pared to small -diam-eter(<4cm)tumors.The overall survival(OS)of bulky barrelshaped tumors was significan tly worse(P<10 -4 ).The same was found for disease -free s urvival(DFS).Conclusion.Bulky exophytic cervic al cancer has an iden-tical surgical morbidity,overall a nd disease -free survival as compared to nonbulky(<4cm)cervical cancer.In view of these identical characteris tics,primary surgical treatment should be considered for p atients with bulky ex-ophytic cervical cancer.
文摘The second mitochondria-derived activator of caspases (Smac) is a novelproapoptotic gene, which plays an important role in the apoptosis-inducing effects of irradiation ontumor cells. The purpose of this study was to investigate the effects of extrinsic Smac genetransfer and its over-expression in radiotherapeutic sensitivities of cervical cancer cells. Afterthe Smac gene was transferred into the cervical cancer cell line HeLa, subcloned cells were obtainedby persistent G418 selection. Cellular Smac gene expression was detected by RT-PCR and Westernblot, while in vitro cell viabilities were detected by trypan blue staining assay. After treatmentwith X-ray irradiation, cellular radiotherapeutic sensitivities were investigated by tetrazoliumbromide colorimetry. Cellular apoptosis and its rate were determined by electronic microscopy,annexin V-FITC and propidium iodide staining flow cytometry. The expression and activities ofcellular caspase-3 were assayed by Western blot and colorimetry. Smac mRNA and protein levels inHeLa/Smac cells and the selected subclone cell line of cervical cancer were significantly higherthan those of HeLa (P <0. 01). There was no significant difference in cellular viabilities betweenthem (P > 0. 05 ) . However, after irradiation with 8 Gy X-ray, growth activities of HeLa/ Smac werereduced by 22.42% (P < 0. 01). When compared with those of HeLa, partial HeLa/Smac cells presentedcharacteristic morphological changes of apoptosis under electronic microscope, with higher apoptosisrates (16. 4% vs. 6. 2% , P < 0. 01 ) ; the caspase-3 expression levels in HeLa/Smac cells wereimproved significantly (P <0. 01) , while its activities were increased by 3. 42 times (P <0. 01).Stable transfer of the extrinsic Smac gene and its over-expression in cervical cancer cell linecould significantly enhance the expression and activities of cellular caspase-3 and ameliorateapoptosis-inducing effects of irradiation on cancer cells, which was a novel strategy to improveradiotherapeutic effects on cervical cancer.
