宫颈癌放疗前应用双氧水阴道冲洗的近期疗效观察

目的观察双氧水在宫颈癌放疗前应用的临床效果。方法将2005年3月~2007年3月在我院行根治性放疗的120例中晚期宫颈癌患者随机分为两组。研究组60例,放疗前常规应用双氧水阴道冲洗;对照组60例,放疗前常规应用新洁尔灭液冲洗。两组患者放射治疗方案相同。比较两组患者局部肿瘤消退情况、治疗效果及并发症发生情况。结果宫颈局部肿瘤缩小50%与100%的肿瘤放射剂量：研究组分别为（20.64±5.03）Gy（、33.09±7.86）Gy;对照组分别为（42.05±14.06）Gy、（50.08±21.02）Gy。肿瘤完全消退时间：研究组为（31.26±10.25）d,对照组为（42.80±9.06）d,两组比较差异有显著性（P〈0.01）。近期有效率、局部复发率、远处转移率及放射性不良反应发生率两组比较差异均无显著性（P〉0.05）。结论宫颈癌患者放疗前常规应用双氧水阴道冲洗能加快肿瘤消退,能否减少放射剂量及是否提高远期疗效有待进一步观察。

Influences of Melatonin on the Growth of HeLa Cells

Aim To investigate the influences of melatonin (MT) on the growth of HeLa cells in vitro. Methods Theantiprolfferation activities of MT were evaluated in HeLa cells by means of trypan blue dye exclusion and MTT vital staining.The morphological changes of HeLa cells induced by MT were observed under transmission electronic microscope. Cell divisioncycle influenced by MT was assessed by a flow cytometry. Results MT produced a certain inhibition of HeLa cells at the con-centration of 2 mmol@ L-1 and prolonged the TD. The fraction of cells inhibited was 61.0%. The IC. so of HeLa cells exposed toMT for 96 h was 2.039 mmol@ L- 1. The flow cytometric analyses showed that exposure to MT for 72 h reduced the number ofHeLa cells in phase S. Under electronic microscope, the HeLa cells exposed to MT for 72 h displayed morphological changesof necrosis, apoptosis, more hetero-chromosome and less somatic chromosome. Conclusion MT showed certain influences onthe growth of HeLa cells. Its mechanism may probably be attributable to reduction of the number of cells in phase S.

Antitumor Actinity of Puqietinone,a Novel Alkaloid fromthe Bulbs of Fritillaria puqiensis

从蒲圻贝母中分得的一种新生物碱蒲贝酮碱，按一定剂量灌胃给药，显示了强的抗小鼠艾氏腹水癌（ＥＡＳ，实体型），宫颈癌（Ｕ＿（１４），实体型）及肝癌（ＨｅＰＡ，实体型）的活性。

Peutz-Jeghers syndrome with small intestinal malignancy and cervical carcinoma

We report a case of 30-year-old woman with Peutz- Jeghers syndrome (PJS). Because of small intestinal obstruction, she received the small intestinal polypectomy in 2001, and the pathological diagnosis was Peutz-Jeghers polyp canceration (mucinous adenocarcinoma, infiltrating full-thickness of the intestine). The patient did not feel uncomfortable after 6 mo of chemotherapy and other management. We kept a follow-up study on her and found that she suffered from cervical cancer in 2007, with a pathological diagnosis of cervical adenosquamous carcinoma.The patient presented with typical features of PJS, but without a family history. The PJS accompanied with both small intestinal and cervical malignancies has not been reported so far in the world.

RESPONSE OF EARLY STAGE BULKY CERVICAL SQUAMOUS CARCINOMA TO PREOPERATIVE ADJUVANT CHEMOTHERAPY

Objective To investigate the potential role of preoperative adjuvant chemotherapy on early stage cervical squamous carcinoma with bulky tumor. Methods One hundred and forty-five patients with cervical squamous cancer stagesⅠb-Ⅱa were investigated, among which17 patients with bulky tumors (≥4 cm) were managed by cisplatin-based chemotherapy for 1-2 courses followed by radical hysterectomy and pelvic lymphadenectomy (BC group). The change of tumor size, pelvic lymph nodes metastasis, cervical wall invasion, the involvement of surgical specimen margin, and the blood loss during operation were assessed after opera-tion and compared with those in 51 patients with bulky tumors (BN group) and 77 patients with small local tumors (S group) who underwent surgery directly. Results (1) The tumor size of 17 patients in BC group were decreased in various degrees after chemotherapy, with 13 pati-ents of clinical effectiveness (76.47%). And the responsiveness pertained to neither histological differentiation nor size of local tumors. (2) Post-operative histology has showed that patients in BC and BN group have higher incidence of lymph node metastasis and deep cervical infiltration (5/68 and 3/68, respectively) than in S group (1/77 and 1/77, respectively) while with no statistical significance. (3) Blood loss during operation in BC group was less than BN and S group. (4) Seventeen patients, including those underwent surgeries of vaginal prolongation and/or ovarian transposition, appeared disease-free survival within the follow-up time. Conclusions Most of patients with bulky early stage cervical squamous carcinoma are sensitive to cisplatin-based chem-otherapy, which could greatly reduce local tumor size and in turn facilitate the following operation by well controlling blood loss.

Expression of Bmi-1,P16,and CD44v6 in Uterine Cervical Carcinoma and Its Clinical Significance

Objective Bmi-1, a putative proto-oncogene, is a core member of the polycomb gene family, which is expressed in many human tumors. The p16 protein negatively regulated cell proliferation, whereas CD44v6 is associated with proliferation as an important protein. Additionally, CD44v6 is an important nuclear antigen closely correlated to tumor metastasis. Tlle present study aims to investigate the expression and significance of Bmi-1, p16, and CD44v6 in uterine cervical carcinoma （UCC）. Methods A total of 62 UCC, 30 cervical neoplasic, and 20 normal cervical mucosal tissues were used ill the current study. The expression of Bmi-1, p16, and CD44v6 in these tissues was determined using immunohistochemical assay. The relationships among the expression of these indices, the clinicopathologic features of UCC, and the survival rate of UCC patients were also discussed. The correlation between Bmi-1 protein expression and p16 or CD44v6 protein in UCC was analyzed. Results The expression of Bmi-l, p16, and CD44v6 was significantly high in cervical carcinoma compared with that in tlle cervical neoplasia and normal colorectal mucosa （P〈0.05）. The over-expression of Bmi-1 protein in UCC was apparently related to the distant metastasis （P〈0.01） and the tumor, nodes and metastasis-classification, i.e. the TNM staging, World Health Organization （P〈0.05）. Nevertheless, the positive expression of p16 protein in UCC was not significantly associated with the clinicopathologic features （P〉0.05）. The Kaplan-Meier survival analysis showed that the over-expression of Bmi-1 significantly decreased the survival rate of UCC patients （P〈0.05）. A strong correlation indicated that there was statistical significance between the expression of Bmi-1 and CD44V6 proteins in UCC （r=0.419, P=0.001）. Conclusions The over-expression of Bmi-1 and CD44v6 protein closely correlate to the tumorigenesis, metastasis, and prognosis of UCC. Bmi-I and CD44v6 may be used to predict the prognosis of cervical carcinoma. Bmi-1 may indirectly regulate the expression of CD44v6 in UCC patients. The positive expression of p16 protein is possibly associated with the tumorigenesis, but not with the metastasis or prognosis of UCC.

Mechanism of anti-tumor effect of HIF-1α silencing on cervical cancer in nude mice

Objective： The aim of our study was to observe the anti-tumor effect of silencing the expression of HIF-1α on cervical cancer in nude mice and to explore its mechanism of action. Methods： Human cervical cancer cell line Siha cells were divided into 3 groups： mock control group, control group transfected with scrambled sequence plasmid, and experimental group transfected with pU-HIF-la-shRNA eukaryotic expression plasmid. Cultured cells of the three groups were inoculated in nude mice to establish cervical cancer-bearing nude mice. HIF-la RNAi assay was performed to evaluate the tumor-suppressive effect of HIF-1α silencing on cervical cancer-bearing nude mice. Immunohistochemistry and Western blot were used to observe the distribution and protein expression of HIF-1α and GLUT1, while RT-PCR was adopted to detect the gene expression of HIF-1α, GLUTI and HK I1. The product of glycolysis （tactic acid） and apoptosis in tumor cells were detected by colorimetry and semi-quantitative TUNEL staining, respectively. Results： The tumor growth in experimental group was significantly slower than that in the two control groups （P 〈 0.05）. In the 50th day after transplantation, the tumor weight in the experimental group was （1.90 ± 0.28） g, significantly lower than （2.95 ± 0.77） g in the control group and （2.54 ± 0.56） g in the mock group （P 〈 0.01）. In the experimental group, the gene and protein levels of HIF-1α were 0.45 ± 0.04 and 1.25 ± 0.92, and the levels of GLUT1 were 0.32 ± 0.02 and 1.25 ± 0.48, respectively. Both indicators in HIF-la and GLUT1 were lower than that in the two control groups （P 〈 0.05）. The expression levels of HK Ⅱ gene and lactic acid in the experimental group were lower than that in the two control groups （P 〈 0.05）, but the apoptotic cells were much more numerous in the experimental group than that in matched control groups （P 〈 0.01）. Conclusion： The 9ene therapy by siRNAtargeted silencing of HIF-1α may down-regulate its downstream genes GLUT1 and HK Ⅱ expression, therefore, to reduce the tumor glycolysis activity and promote tumor cell apoptosis, and exert a tumor-suppressing effect in vivo.

Effects of trichostatin A(TSA)on growth and gene expression in HeLa cells

Objective： To investigate the expressions of p53, RB1, Fas, c-fos, Ras, EGFR mRNA in human cervical cancer （HeLa） cell in response to the trichostatin A （TSA）. Methods： We took count of HeLa cells in different incubation times with TSA （0.2μm/L）. The result indicated that HeLa cells changed evidently when HeLa cells were incubated for 36 h. Then, we investigated the genes expression （mRNA levels） of HeLa cells after treatment for 36 h using SYBR green real-time PCR. Results： We demonstrated that trichostatin A （TSA） could make human cervical cancer （HeLa） cell morphological change and induce HeLa cell apoptosis. Furthermore, the data suggest that TSA-induced down-regulation of p53, RB1, Fas, but upregulated c-fos gene expression after treatment for 36 h, and Ras, EGFR did not show obvious response to TSA treatments. Conclusion： TSA has different effects on gene expression.

Effects of the anti-tumor composition ofthe acetoacetate extract of vitex negundoseed on the growth of human cervical carcinomaxenografts in nude mice

Objective： The aim of our study was to investigate the effects of the anti-tumor composition of the acetoacetate extract of Vitex Negundo Seed （EVn-50） on the growth of human cervical carcinoma HeLa cells xenografts in nude mica and its possible molecular mechanism. Methods： Models of human cervical cancer HeLa cells xenografts transplanted subcuta- neously in nude mice were established and randomly divided into 7 groups （each group including 5 nude mice）： saline group, Taxol group, EVn-50 group, comp-6 group, comp-7 group, comp-8 group and comp-10 group. The volume and weight of Xe- nograts were observed and compared. The alteration of the weight of nude mice, and the change of serum levels ofLDH, ALT, Cr and WBC counts were examined and compared. The apoptotic rate of human cervical carcinoma HeLa cells xenografts was analyzed by FCM. The expressions of P53 and Bcl-2 proteins of HeLa cells xenografts were determined by Western blot- ting. Results： EVn-50 and its fractionated extracts could significantly suppress the increasing volume and weight of human cervical carcinoma HeLa cells xenografts in nude mice models in time-dependent manner, yet had no significant effect on the weight of nude mice, the serum levels of LDH, ALT, Cr and WBC were counted. When the xenografts were treated with EVn-50 and its fractionated extracts for 16 days, the apoptotic rate of xenografts cells were significantly increased, and the expression of P53 protein was up-regulated and protein level of Bcl-2 was decreased. Conclusion： EVn-50 and its fractionated extracts could suppress the growth of human cervical carcinoma HeLa cells xenografts in nude mice, which may be related to its pro- motion on xenografts cells apoptosis through down-regulation of Bcl-2 expressionPand activation of P53 expression.

Aberrant Expression of Notch1 in Cervical Cancer

OBJECTIVE To investigate the putative role of the Notch1 receptor in cervical cancer carcinogenesis and progression. METHODS The expression of the Notch1 protein was analyzed by a Western-blotting approach in 40 cervical cancer and 30 normal cervical tissues. Some tissues were examined using RT-PCR to determine mRNA levels. Celluar localization of the Notch1 protein in the paraffin-embedded cervical tissues was also analyzed by immunohistochemistry. RESULTS The Notch1 protein was detected in all 30 normal cervical tissues. In contrast, only 6 samples of 40 cervical cancer tissues showed Notch1 expression. The level of the Notch1 protein expression was significantly lower in cervical cancer tissues than that in normal tissue samples. In agreement with these observations, levels of Notch1 mRNA were found to be substantially down-regulated in cervical cancer tissues. In the immunohistochemistry staining assay, the Notch1 protein was shown to localize predominantly in the cytoplasm and nucleoli of the normal cervical squamous epithelium of the cervix, but no staining was observed in the cervical cancer cells. Notch1 expression was observed to correlate with the clinical disease stage, but there were no correlations with age, tumor size, grade or lymph node metastasis （P〉0.05）. The levels of Notch1 protein expression were significantly higher in early stages （Ⅰ-Ⅱa, 66.7%） compared to those in the advanced stages （Ⅱb～Ⅳ,12.6%）（P=0.001）. CONCLUSION Notch1 may play a role as a tumor suppressor in cervical tumorigenesis. Determination of Notch1 expression may be helpful for preoperative diagnosis and accuracy of staging. But its clinical use for cervical cancer requires further investigation.

Pharmacokinetic comparison between ultraselection of uterine artery and peripheral vein chemotherapy of carboplatin in cervical cancer

Objective:The present study is to compare pharmacokinetics difference of carboplatin by using ultraselection uterine artery with by using peripheral vein in cervical cancer.Methods:Thirteen patients with locally advanced cervical cancer who had been proved by pathobiology were randomly divided into two groups:the ultraselection uterine artery group(group A, n = 6) and the peripheral vein(group B, n = 7).Carboplatin was administered by infusing into artery or vein in both groups at the dosage of 300 mg/m2.Tissues from the cervical tumor were collected at different times after infusion in both groups and then analyzed.Results:The peak concentration of platinum in tumor tissue was about 2.79 times higher in group A than that of group B(P < 0.05).The platinum concentrations in tumor in group A reached its peak levels immediately after infusion.But, group B had delayed time.While, for the time point of 0 min, when the administration finished immediately, the platinum concentration in tumor was significantly higher when compared with group B(P < 0.05).The tumor tissue area under the concentration(AUC) of carboplatin was about 2 times higher in group A than that of group B(P < 0.05).Conclusion:We observed the pharmacological advantages of chemotherapy by using ultraselection uterine artery administration of chemotherapeutic agent carboplatin in tumor tissue which provided theoretical basis and laboratory parameters of the intra-arterial chemotherapy for gynecologic malignancy.