Effect of autophagy gene Beclin 1 on the growth of cervical cancer HeLa cells in vitro and vivo / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the effects of autophagy gene Beclin 1 on growth of cervical cancer HeLa cells in vitro and vivo.</p><p><b>METHODS</b>The eukaryotic expression vector of Beclin1 was constructed and transfected via lipofectamine into HeLa cells. The experimental cells were classified into 3 groups: pcDNA3.1(+)-Beclin1 group,pcDNA3.1(+) group and HeLa group. Real time-ploymerase chain reaction and Western blot were used for detecting expression of Beclin1 mRNA and protein in the transfected cells. Flow cytometry (FCM) was employed to observe the effect of transfection on the apoptosis of HeLa cells, and proliferation was analyzed by MTT assay. The formation of autophagic vacuoles was measured by MDC staining. HeLa cells transfected with plasmid pcDNA3.1(+)-Beclin1 and pcDNA3.1(+) were inoculated subcutaneously in nude mice. The carcinogenic and growth activities of cancer cells in vivo were observed.</p><p><b>RESULTS</b>Eukaryotic expression vector pcDNA3.1(+)-Beclin1 was constructed successfully. It significantly improved the expression of Beclin1 mRNA and protein in HeLa cells. The proliferation of HeLa cells was inhibited, and the inhibition rate was 58.7%. FCM investigation showed that the apoptotic rate was (28.22 ± 2.34)% of pcDNA3.1(+)-Beclin1 group, significantly higher than the (14.6 ± 4.6)% in the pcDNA3.1(+) group and (11.2 ± 3.0)% in the HeLa group (P < 0.05). The monodansylcadaverin (MDC) staining showed significantly more autophagic vacuoles in the pcDNA3.1(+)-Beclin1 group (10.9%) than that in the pcDNA3.1(+) group (3.1%) and HeLa group (2.5%) (P < 0.05). After transfected with vector pcDNA3.1(+)-Beclin1, the carcinogenic activity of HeLa cells was decreased in nude mice, and the inhibition rate of tumor growth was 52.2%.</p><p><b>CONCLUSIONS</b>Autophagy gene Beclin 1 overexpression can inhibit the proliferation and growth of HeLa cells in vitro and vivo,while promote autophagy and apoptosis of HeLa cells. So it might be one of new gene therapy strategies for cervical carcinoma.</p>

Growth inhibitory effects of Beclin1 gene on epithelial ovarian cancer SKOV3 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the effect of Beclin1 overexpression on the growth of ovarian carcinoma cell line SKOV3 in vitro and in vivo.</p><p><b>METHODS</b>The recombinant plasmid pcDNA3.1/Beclin1 was constructed and transfected into SKOV3 cells via lipofectamine 2000. MTT assay was used to evaluate the effect of Beclin1 overexpression on the proliferation and growth of the transfected cells, whose apoptosis and autophagy were analyzed by flow cytometry. SKOV3 cells transfected with the plasmids pcDNA3.1/Beclin1 or pcDNA3.1 were inoculated subcutaneously in nude mice, and their carcinogenic and growth activities in vivo were evaluated.</p><p><b>RESULTS</b>MTT assay showed that transfection with pcDNA3.1/Beclin1 significantly inhibited the proliferations of SKOV3 cells, with a cell inhibition rate of 58.68% (P<0.05). The transfection also resulted in a cell apoptosis rate of (21.26-/+3.89)%, significantly higher than that of pcDNA3.1 trasnfection (P<0.05). Flow cytomerty showed that pcDNA3.1/Beclin1 transfection of SKOV3 cells produced a significantly higher MDC fluorescent intensity than pcDNA3.1 transfection. The SKOV3 cells transfected with vector pcDNA3.1/Beclin1 also showed decreased carcinogenic activity in nude mice, with a growth inhibition rate of 50.27%.</p><p><b>CONCLUSION</b>Beclin1 overexpression can inhibit the proliferation and growth of SKOV3 cells in vitro and vivo, suggesting its potential role in gene therapy of ovarian carcinoma.</p>

Malignant peripheral nerve sheath tumor of the vagina

Malignant peripheral nerve sheath tumors [MPNSTs] usually develop in major nerve trunks, giving rise to tumors in the proximal portions of the upper and lower extremities and trunk. The MPNSTs primarily occurring in the female genital tract are quite rare. We describe a case of MPNST of the vagina and discuss its diagnosis and treatment. This patient was treated definitively with radical local excision and is well with no signs of recurrence 2 years postoperatively

Diagnosis and management of 57 cases with rectovaginal endometriosis / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To study the diagnosis and therapy of the rectovaginal endometriosis.</p><p><b>METHODS</b>Clinical data of 57 women with rectovaginal endometriosis admitted to the West China Second University Hospital of Sichuan University in last two years,were retrospectively reviewed.</p><p><b>RESULT</b>The average age of patients was 40.1 years. The main clinical manifestations were dysmenorrheal, changes of menorrhea and digestive stimulation. The diameter of deep endometriosis nodules was between 1-6 cm, and 77% were found to have more than one nodules. Seven of these patients had positive results in transvaginal ultrasonography; 61%(11/18) patients had elevated CA125 levels. Thirteen patients were given preoperational medical treatment, but had no effect. All patients, except one accepted laparotomic therapy of complete excision of endometriosis nodules; 23 cases underwent drug therapy after operation. No patients had recurrence in recto-vaginal septum after complete excision; only one recurred in right ovary. Patients who failed to remove the total lesion showed improvement in pain.</p><p><b>CONCLUSION</b>Diagnosis of the rectovaginal endometriosis is based on symptoms, vaginal and rectal examination, and auxiliary examination. Complete excision of endometriosis nodules is the main therapeutic method.</p>

Expression of THY1 gene in epithelial ovarian cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To detect the expession of THY1 in ovarian serous cystadenocarcinoma tissues.</p><p><b>METHODS</b>Immunohistochemistry was performed to detect the expression of THY1 gene in formalin-fixed, paraffin-embedded specimens of normal ovaries (n = 25), ovarian serous cystadenoma (n = 25), and serous cystadenocarcinoma (n = 53). The correlation of THY1 expression with clinicopathological parameters was statistically analyzed.</p><p><b>RESULTS</b>The positive expression rates of THY1 protein in normal ovaries, ovarian serous cystadenomas and ovarian serous cystadenocarcinomas were 60.0% (15/25), 72.0% (18/25) and 34.0% (18/53), respectively. The values of IOD of THY1 protein expression were 288,449.2 +/- 60,087.3, 271,655.6 +/- 66,588.7 and 252,087.6 +/- 45,559.4, respectively. The expression of THY1 protein was significantly down-regulated in ovarian serous cystadenocarcinoma tissues compared with that in normal ovarian tissues and ovarian serous cystadenoma tissues (P < 0.05). THY1 expression was negatively correlated with surgical-pathological staging, histological differentiation and lymph node involvement (P < 0.05).</p><p><b>CONCLUSION</b>The decreased level of THY1 expression may be related with the occurrence and development of ovarian serous cystadenocarcinoma.</p>

Inhibiting effect of HPV16 E6 small interfering RNA on the growth of human cervical carcinoma xenotransplanted in nude mice / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the inhibiting effect of human papillomavirus 16 E6-specific small interfering RNA (siRNA) on cervical cancer transplanted in nude mice.</p><p><b>METHODS</b>CaSki cells transfected with HPV16 E6 siRNA were transplanted into nude mice. HPV16 E6 siRNA was injected into the tumor, and the control group was treated with NS. Tumor growth was monitored once every other day. Terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL) was performed to detect apoptosis. The expression of E6 and p53 protein was assessed by immunohistochemistry. The contents of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured and the liver and kidney were examined by histopathology.</p><p><b>RESULTS</b>Inhibition of tumor growth was demounstrated after treatment with HPV16 E6 siRNA. The volume and weight of tumors were significantly decreased in comparison with those of control group (P < 0.05). Apoptosis occurred more frequently in the experiment group than in the control. The expression of E6 and p53 was down-regulated. The contents of ALT and AST underwent no significant changes, and the histopathology of liver and kidney showed no abnormal changes.</p><p><b>CONCLUSION</b>The growth ability of human cervical cancer xenograft tumors in nude mice can be inhibited by HPV16 E6-specific siRNA, with no toxic side effect on the liver. It may provide an useful method of gene-therapy against cervical cancer.</p>

Growth inhibitory effects of THY1 gene on epithelial ovarian cancer SKOV3 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct THY1 eukaryotic expression plasmid and study its effects on the growth of epithelial ovarian cancer cell line SKOV3.</p><p><b>METHODS</b>THY1 gene fragment was obtained from normal human ovarian tissue using RT-PCR, and inserted into the eukaryotic expression plasmid pcDNA3.1(+) to construct the recombinant plasmid pcDNA3.1(+)-THY1, which was transformed into E. coli JM109 followed by selection of the positive clones containing the target inserts. The eukaryotic expression plasmid was analyzed by PCR, restriction endonucleases digestion and DNA sequencing. SKOV3 cells divided into SKOV3-THY1, SKOV-3-Null and SKOV3 groups were transfected via liposome with the recombinant plasmid pcDNA3.1(+)-THY1, empty plasmid, or not transfected, respectively. The expression of THY1 mRNA and its protein were examined by RT-PCR and Western blot methods. The cell growth and apoptosis were evaluated by MTT assay and flow cytometry.</p><p><b>RESULTS</b>The gene fragment of exogenous THY1 was correctly inserted into the eukaryotic expression plasmid pcDNA3.1(+) as verified by PCR, restriction endonucleases digestion and DNA sequencing, and recombinant expression plasmid pcDNA3.1(+)-THY1 transfection resulted in stable expression in SKOV3 cells as shown by RT-PCR and Western blotting. The cell growth inhibition rate of SKOV3-THY1 group (56.6% at the fifth day) was significantly higher than that of the SKOV3-Null group (12.5%, P<0.05), and the cell apoptosis rate in SKOV3-THY1 group (31.8%) was significantly higher than those in SKOV3-Null group (10.5%) and SKOV3 group (9.8%, P<0.05), but the apoptosis rate between the latter two groups was similar (P>0.05).</p><p><b>CONCLUSIONS</b>The recombinant plasmid pcDNA3.1(+)-THY1 can be expressed stably in human ovarian cancer cell line SKOV3. THY1 transfection can inhibit the growth of SKOV3 cells in vitro, suggesting the important role of THY1 gene in pathogenesis and development of ovarian cancer.</p>

The relationship between the implantation of ovarian stimulated embryos and the expression of leukemia inhibitory factor mRNA in mice metrium / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To check the expression of leukemia inhibitory factor (Lif) mRNA, and to study the impact of ovarian stimulation on the ability of embryo implantation in mice.</p><p><b>METHODS</b>Pregnancy models of mice were established. The relationship between the implantation of ovarian stimulated embryos and the expression of Lif mRNA in mice metrium was analyzed.</p><p><b>RESULTS</b>The group of recipients which the transfered embryos were from stimulated cycles had lower pregnancy and implantation rate compared with the group of recipients which the transfered embryos were from non-stimulated cycles (20.00%, 8.33% vs 55.00%, 35.00%). The Lif mRNA expression was similar in the groups of recipients which the transfered embryos were from stimulated and non-stimulated cycles, so was in the groups of recipients which had single or more than one baby, but higher in the group of pregnancy recipients than in the group of unpregnancy recipients.</p><p><b>CONCLUSION</b>Ovarian stimulation may reduce the ability of embryo implantation in mice. Lif mRNA expression is related to the implantation, but not parallel to the number of implantation.</p>

The effect of basic fibroblast growth factor in ovarian cancer growth and angiogenesis / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate whether basic fibroblast growth factor (bFGF) can induce the proliferation, invasion and angiogenesis of ovarian cancer or not.</p><p><b>METHODS</b>Human ovarian cancer cell lines SKOV(3) 1 x 10(4)/ml were plated in 24-well dishes, bFGF at 5, 10,15 and 20 ng/ml was added and crystal violet staining was given daily for 8 days, cell numbers were counted by determining OD490. SKOV(3) cells were plated in the center of 50% extra cellular matrix gel, bFGF at 5 and 10 ng/ml was added and the migration distance of cells was measured daily. SKOV(3) 5 x 10(7)/ml were transplanted to BALB/c nude mice subcutaneous. One week later, bFGF, bFGF-MAb or 0.9% nature sodium was injected subcutaneously surrounding the tumor twice a week. Eight weeks later, the experiment ended and the volume of the tumors were measured. Intratumoral microvessel density (MVD) was measured by immunohistochemistry staining for factor VIII.</p><p><b>RESULTS</b>bFGF at 0-10 ng/ml could stimulate the proliferation of SKOV(3) concentration dependently (P<0.05). On the fifth day, the cell proliferation in 10 ng/ml group was 121% above control. bFGF could stimulate the invasion of SKOV(3) concentration dependently (P<0.05). On the seventh day, the migration distance in 5 ng/ml group was 1.16 cm and 153% above control, and that in 10 ng/ml group was 1.86 cm and 245% above control. The average volume of transplanted tumors and MVD in bFGF group were 180% and 146% above control respectively those in bFGF-MAb group were 63.7% and 62.8% above control respectively.</p><p><b>CONCLUSION</b>bFGF can stimulate proliferation, invasion and angiogenesis of ovarian cancer markedly; bFGF-MAb can inhibit the angiogenesis and growth of ovarian cancer.</p>

Tissue microarray and in situ hybridization study on hypoxia inducible factor 1alpha mRNA expression in epithelial ovarian tumor / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the expression and significance of hypoxia inducible factor 1alpha (HIF-1alpha) in epithelial ovarian tumors.</p><p><b>METHODS</b>The expression of HIF-1alpha mRNA in 295 patients with epithelial ovarian tumor was analyzed retrospectively by high-throughput tissue microarray and in situ hybridization, which was compared with 13 normal ovarian tissue samples.</p><p><b>RESULTS</b>The expression rates of HIF-1alpha mRNA were 0, 13.2%, 42.1% and 81.9% in normal ovarian tissue, benign, borderline and malignant ovarian tumors. Expression rate of HIF-1alpha mRNA in borderline and invasive tumor was significantly higher than those in normal ovarian tissue and benign tumor (P < 0.001). Statistical analysis revealed that the expression of HIF-1alpha mRNA was not related to FIGO stages or histological subtypes. Close negative relation was observed between the expression of HIF-1alpha mRNA and tumor histological differentiation (P < 0.001).</p><p><b>CONCLUSION</b>The overexpression of HIF-1alpha may play an important role in oncogenesis of epithelial ovarian tumor. Tissue microarray is an efficient technique of molecular biology.</p>

Clinicopathologic and immunohistochemical study of primary non-Hodgkin lymphoma of the female genital system / 中华妇产科杂志

Objective To investigate the clinicopathology and immunophenotype of primary non- Hodgkin lymphoma(NHL)of the female genital system,and to analyze the prognosis of such tumors. Methods Clinicopathologic features of 43 cases of primary NHL of the female genital system were studied retrospectively,with the histological classification based on the Classification of Haematopoietic and Lymphoid Tumors(WHO,2001).Immunochemistry technique,in-situ-hybridization and polymerase chain reaction methods were used to detect the immunophenotype,epstein barrvirus(EB)virus infection status and immunoglobulin heavy chain gene rearrangement,respectively.Results(1)Primary lesions:there were 24 cases of lymphoma originating in the ovary,3 cases in the endometrium,10 cases in the cervix,2 cases in the vagina and 4 cases in the vulva.(2)Staging:12 cases(28%)were in stage Ⅰ,9 cases (21%)in stage Ⅱ,and 22 cases(51%)in stage Ⅲ.(3)Histological classification:37 cases(86%)were diffuse large B cell lymphoma(DLBCL),3 cases were Burkitt lymphoma and the remaining 3 cases were unspecified peripheral T-cell lymphoma according to biopsy,immunophenotype analysis,in-situ- hybridization technique and IgH gene rearrangement detection.(4)Prognosis analysis:increase in the level of lactic acid dehydrogenase,stage Ⅲ,DLBCL and single operation suggest poor prognosis.Conclusions Establishment of the diagnosis of primary NHL of the female genital system is based on biopsy, immunophenotype analysis,in-situ-hybridization technique and IgH gene rearrangement detection,which play important roles in diagnosis and differential diagnosis of the tumor.Combined therapy is the first choice of therapeutic regimens.