The effect of edaravone combined with DL-3-N-butylphthalide on the levels of tumor necrosis factor-alpha, interleukin-10, neuron-specific enolase and effect in patients with acute cerebral infarction.

This study observed the effects of edaravone combined with Dl-3-N-butylphthalide (NBP) on the serum levels of tumor necrosis factor-alpha (TNF-α), interleukin-10 (IL-10), and neuron-specific enolase (NSE), and its therapeutic effect in patients with acute cerebral infarction (ACI). The purpose of this study was to explore whether edaravone combined with NBP could improve the neurological function of patients with ACI. A total of 86 patients with ACI were enrolled in this study; 43 patients were randomly assigned to the control group and treated with edaravone only, while the other 43 patients were assigned to the intervention group and treated with a combination of edaravone and NBP. The course of treatment lasted 14 days, and the basic drug treatment was the same in both groups. The effective rate of activity of daily living scores (ADL) was significantly higher in the intervention than in the control group, and the difference was statistically significant (P<0.05). After the treatment had been administered, the National Institute of Health Stroke Scale scores of the two groups were lower than before the treatment, and the scores were lower in the intervention compared with the control group; the difference was statistically significant (P<0.05). After the treatment had been administered, the serum levels of TNF-α and NSE were significantly lower in the intervention than in the control group, and the serum IL-10 level was significantly higher in the intervention than in the control group; the differences were statistically significant (P<0.05). Edaravone combined with NBP improved the neurological function of patients with ACI, improved their quality of life, significantly decreased the serum levels of TNF-a and NSE, increased the serum IL-10 level, and had a better effect. This combination therapy method can be adopted in clinics to treat patients with ACI.

The Carbon Sink Potential of Southern China After Two Decades of Afforestation.

Afforestation and land use changes that sequester carbon from the atmosphere in the form of woody biomass have turned southern China into one of the largest carbon sinks globally, which contributes to mitigating climate change. However, forest growth saturation and available land that can be forested limit the longevity of this carbon sink, and while a plethora of studies have quantified vegetation changes over the last decades, the remaining carbon sink potential of this area is currently unknown. Here, we train a model with multiple predictors characterizing the heterogeneous landscapes of southern China and predict the biomass carbon carrying capacity of the region for 2002-2017. We compare observed and predicted biomass carbon density and find that during about two decades of afforestation, 2.34 PgC have been sequestered between 2002 and 2017, and a total of 5.32 Pg carbon can potentially still be sequestrated. This means that the region has reached 73% of its aboveground biomass carbon carrying capacity in 2017, which is 12% more than in 2002, equal to a decrease of 0.77% per year. We identify potential afforestation areas that can still sequester 2.39 PgC, while old and new forests have reached 87% of their potential with 1.85 PgC remaining. Our work locates areas where vegetation has not yet reached its full potential but also shows that afforestation is not a long-term solution for climate change mitigation.

[An experiment control study on the ovarian reserve function after cisplatin intraperitoneal or intravenous chemotherapy in rats model].

Objective: To compare the impact on the ovarian reserve function after cisplatin intraperitoneal or intravenous chemotherapy in rats model. Methods: Thirty 8-weeks old female Sprague Dawley rats were randomly assigned to control group (group A, n=10), intraperitoneal chemotherapy group (group B, n=10) and intravenous chemotherapy group (group C, n=10). Cisplatin was diluted by normal saline (NS) into 4 mg/ml. On the first day of chemotherapy, 0.2 ml cisplatin dilution was injected into the abdomen of rats in group B, isodose cisplatin was injected into vein and 1.8 ml NS was injected into abdomen of rats in group C, 2.0 ml NS was injected into abdomen of rats in group A for control. Feed the three groups rats and test the anti-Mullerian hormone (AMH) in serum on day 0 (just before injection), day 10 and day 20 by ELISA, count the numble of follicle in bilateral ovaries on day 20. Results: (1) The levels of serum AMH in the three groups before and after chemotherapy were compared: ① comparison between groups: On day 10 and day 20, the AMH level in group B [(64.5±2.9), (68.6±3.4) ng/L] and group C [(76.1±4.9), (91.3±3.9) ng/L] was significantly lower than that in group A [(120.1±5.3), (121.7±4.6) ng/L; P<0.01], AMH level in group B was significantly also lower than that in group C (P=0.000). ② Comparison within groups: the AMH level on day 0 was significantly lower than that on day 10 and day 20 in group A (P<0.01), but there was no significant difference between day 10 and day 20 (P=0.427). The AMH level on day 0 was significantly higher than those on day 10 and day 20 in group B (P<0.01) and group C (P<0.01). There was no difference in AMH level between day 10 and day 20 (P=0.124) in group B, but the level was significant lower on day 10 than that on day 20 in group C (P=0.011). (2)Comparison of the number of follicles in ovaries of three groups 20 days after chemotherapy: the follicles number in group A (35±13) was greater than that in group B (16±9, P=0.003) and similar with group C (31±16, P=0.474) on day 20. The follicles number in group B was significantly less than that in group C (P=0.018). Conclusions: In the present study, both intravenous and intraperitoneal chemotherapy have impacts on ovarian reserve function and the latter might be more serious. The level of AMH will rise again over time after chemotherapy and rats undergo intravenous chemotherapy would recover faster. The results suggest that the ovarian reserve function need more time to recover after intraperitoneal chemotherapy. For patients with ovarian cancer who want to preserve fertility function, intravenous chemotherapy might be more appropriate.

Human kallikrein 5 as a novel prognostic biomarker for triple-negative breast cancer: tissue expression analysis and relationship with disease course.

The purposes of this study were to analyze the expression and distribution of human kallikrein 5 (hK5) in triple-negative breast cancer (TNBC) tissues, to establish a standard operating procedure (SOP) for its immunohistochemical assay, and to evaluate the possibility of hK5 being a prognostic biomarker for TNBC. Recombinant hK5 protein and specific antibody were prepared, and the expression and distribution of hK5 in TNBC tissues were detected using immunohistochemistry. An SOP for immunohistochemical staining of hK5 in TNBC tissues was established to allow automatic staining under optimized conditions. The resulting images were digitized for evaluation and statistical analysis via a human scoring system. Our results showed that expression of hK5 protein could predict the progression of TNBC. Pearson's chi-square test results showed that high hK5 expression in tumor stromal cells was significantly correlated with distal metastasis (P = 0.039). A high staining score for lymphocyte infiltration in tumor stroma was significantly correlated with low histological grade of tumor (P = 0.025). Univariate and multivariate Cox regression analyses verified that the staining score for hK5 in tumor stromal cells may be a biomarker for poor prognosis in TNBC patients (univariate HR = 2.289, 95%CI = 1.362-3.848, P = 0.002; multivariate HR = 2.105, 95%CI = 1.189-3.727, P = 0.011). In conclusion, the expression level of hK5 in tumor stromal cells is a promising biomarker for poor prognosis in TNBC. Patients with high histological grade are more prone to distal metastasis and aggressive tumor progression.

Combination of adenovirus-mediated thymidine kinase gene therapy with cytotoxic chemotherapy in bladder cancer in vitro.

We evaluated efficacy, toxicity and potential synergism of adenoviral-mediated thymidine kinase (tk)- ganciclovir (GCV) gene therapy in combination with 4 cytotoxic chemotherapeutic agents (doxorubicin, cisplatin, mitomycin C, and methotrexate) in 3 human bladder cancer cell lines. Cell lines were exposed to (1) 10 different concentrations of adenovirus expressing tk plus GCV; (2) 8 different concentrations of either doxorubicin, methotrexate, mitomycin C or cisplatin; or (3) combination treatment consisting of either low-, medium- or high-dose tk-GCV gene therapy plus 8 different concentrations of a single chemotherapeutic agent. Cell survival was determined using a MTT-based cell proliferation-assay. For most combinations, adding chemotherapy to tk-GCV gene therapy did not result in any therapeutic benefit. In some scenarios, we observed modest improvement with combinations of high-dose tk-GCV gene therapy and high-dose standard chemotherapy over tk-GCV monotherapy. Low concentrations of methotrexate enhanced the antitumor effects of low- and medium-dose tk-GCV gene therapy. Low level negative interference between tk-GCV gene therapy and chemotherapy occurred in some combinations but was overall negligible. In general, adding chemotherapy to tk-GCV gene therapy did not demonstrate significant therapeutic benefit in vitro. High doses of chemotherapeutic agents should be used in combination with tk-GCV gene therapy in order to take advantage of the occasional instance where modest improvement occurred with combination therapy. Additional studies exploring the role of methotrexate in enhancing the tk-GCV system are required. Investigation of other, potentially more synergistic chemotherapeutic agents in combination with tk-GCV is warranted.

Adenovirus-mediated thymidine kinase gene therapy for recurrent ovarian cancer: expression of coxsackie-adenovirus receptor and integrins alphavbeta3 and alphavbeta5.

OBJECTIVE: Ten patients with recurrent ovarian cancer received a combined treatment of optimal tumor debulking, adenovirus-mediated herpes simplex virus thymidine kinase gene therapy (GT), and systemic application of acyclovir or valacyclovir and topotecan. Biopsies were taken at the time of secondary debulking about 1 month after the application of GT and chemotherapy and were analyzed for expression of coxsackie-adenovirus receptor (CAR) and integrins alphavbeta3 and alphavbeta5 with respect to treatment response. METHODS: Treatment modalities and study design have been described recently. Immunohistochemistry was used to visualize expression of CAR and integrins alphavbeta3 and alphavbeta5 in tumor samples taken before and after application of GT. RESULTS: Before GT six of ten patients presented with CAR-positive and four with CAR-negative tumors. After GT all tumors showed CAR expression. Integrin alphavbeta3 was found in all tumors before and after GT. Expression of integrin alphavbeta5 was seen in eight of ten tumor samples before GT and in all samples after GT. CONCLUSION: Despite the importance of CAR and integrin expression for successful adenovirus internalization, other cell surface receptors might be involved in this process. It is too early to decide whether expressions of CAR and integrin alphavbeta3/alphavbeta5 on tumor cells are appropriate additional inclusion criteria for the enrollment of patients in GT trials. Further research is necessary to evaluate the effect of GT plus chemotherapy on CAR and integrin expression.

Histologic and immunohistochemical analysis of tissue response to adenovirus-mediated herpes simplex thymidine kinase gene therapy of ovarian cancer.

Herpes simplex virus (HSV) thymidine kinase (tk) gene incorporated into adenovirus was delivered intraperitoneally (ip) followed by an antiherpetic prodrug and topotecan in patients with recurrent epithelial ovarian cancer. Tissue response was evaluated. Ten patients underwent secondary debulking with subsequent delivery of ADV-HSV-tk therapy. Two patients each were treated at dose level 1 (2 x 10(10) vector particles = VP), 2 (2 x 10(11) VP), and 3 (2 x 10(12) VP); four patients were treated at dose level 4 (2 x 10(13) VP). Five patients underwent second-look surgery about one month after gene therapy (GT). Treatment response, presence of vector DNA, protein expression of steroid hormone receptors, p53, c-erbB2 and Ki67 protein were analyzed. At second-look, two out of five patients were tumor-free and none of their peritoneal biopsies showed vector DNA. After GT, the vital tumor mass was smaller, desmoplastic reaction had increased, and tumors were less differentiated with an increase of Ki67 expression. There was no change in expression of hormone receptors, p53, or c-erbB2. ADV-HSV-tk GT appears to eliminate cells with higher differentiation first and might induce fibrosis. Dedifferentiation might render residual cells more sensitive to chemotherapy secondary to their subsequent higher mitotic activity.

Adenovirus-mediated thymidine kinase gene therapy in combination with topotecan for patients with recurrent ovarian cancer: 2.5-year follow-up.

OBJECTIVE: Patients with recurrent ovarian cancer were treated intraperitoneally (ip) with a replication-deficient recombinant adenovirus (ADV) containing the herpes simplex virus thymidine kinase gene. Vector delivery was followed by intravenous administration of an antiherpetic prodrug and a topoisomerase I inhibitor. METHODS: Ten patients with stage IIIc epithelial ovarian cancer underwent secondary debulking to < or =0.5 cm residual tumor. Patients with normal ip flow received delivery of ip ADV. Two patients each were treated on dose level 1 (2 x 10(10) vector particles), dose level 2 (2 x 10(11)), and dose level 3 (2 x 10(12)); four patients were on dose level 4 (2 x 10(13)). Acyclovir and topotecan were started 24 h after vector delivery. Five patients underwent second-look surgery about 4 weeks after application of gene therapy (GT). RESULTS: At the time of the second-look surgery, two out of five patients were free of tumor. Four weeks after GT none of the peritoneal biopsies showed residual vector DNA. Patients pretreated with an average of three different chemotherapeutic drugs and two different chemotherapy regimens had a median overall survival (OS) of 18.5 months. Three patients are still alive 30, 30, and 31 months after GT. CONCLUSION: With the combination of secondary optimal debulking, GT, and topotecan, median OS was about one-third longer than in previously reported second-and third-line trials. Survival was comparable to that of patients of other studies with secondary cytoreductive surgery in combination with chemotherapy.

Adenovirus-mediated suicide gene therapy for bladder cancer: comparison of the cytomegalovirus- and Rous sarcoma virus-promoter.

BACKGROUND: To compare efficacy and toxicity of the human cytomegalovirus-immediate-early (CMV) promoter and the Rous-sarcoma-virus (RSV) promoter to express thymidine kinase (tk) for adenovirus-mediated suicide gene therapy of experimental bladder cancer in vivo and in vitro. MATERIALS AND METHODS: In vitro: 3 human (5637, RT-4 and TCC-SUP) and one murine (MBT-2) bladder cancer cell line were exposed to ADV/RSV-tk or ADV/CMV-tk vectors and cell survival was determined. In vivo: Subcutaneous tumors were established and adenovirus vectors were injected 10 days later. RESULTS: In vitro: ADV/CMV-tk was up to 4 times more potent in terms of cell killing than ADV/RSV-tk. In vivo: ADV/CMV-tk had a three-fold higher antitumor potency per viral particle as compared to ADV/RSV-tk. Higher doses of ADV/CMV-tk caused treatment-associated hepatotoxicity. CONCLUSIONS: Our results confirm the efficacy of adenovirus-mediated tk suicide gene therapy in the treatment of experimental bladder cancer. Dose-related toxicity was greater with the use of ADV/CMV-tk, but lower doses achieved the same efficacy as ADV/RSV-tk.

Thymidine kinase gene therapy with concomitant topotecan chemotherapy for recurrent ovarian cancer.

INTRODUCTION: Patients with recurrent ovarian cancer were treated with a replication-deficient recombinant adenovirus containing the herpes simplex virus thymidine kinase gene administered intraperitoneally (i.p.) followed by administration of an anti-herpetic prodrug and topotecan. MATERIALS AND METHODS: A total of 10 patients with stage IIIc epithelial ovarian cancer underwent secondary debulking to < or =0.5 cm residual tumor. Patients with normal i.p. flow received i.p. delivery of adenovirus. Two patients each were treated on dose level 1 (2 x 10(10) vector particles (VP)), dose level 2 (2 x 10(11) VP), and dose level 3 (2 x 10(12) VP); four patients were treated on dose level 4 (2 x 10(13) VP). Acyclovir and topotecan were started 24 hours after vector delivery. RESULTS: No patient treated at any dose level incurred unanticipated toxic effects, and all side effects resolved. The most common adverse event was myelosuppression: grade 3 or 4 thrombocytopenia with grade 2-4 anemia in three patients and grade 3 or 4 neutropenia in eight patients. Three patients developed thrombocytosis and three patients had a mild elevation of serum glutamic pyruvic transaminase/alanine aminotransferase. Temperature elevations that were not associated with detectable infection occurred in two patients. DISCUSSION: I.p. delivery of adenoviral vector with concomitant topotecan chemotherapy was well tolerated without significant lasting toxicities. Side effects were independent of the dose of adenoviral vector.

Androgen receptor gene mutations do not occur in ovarian cancer.

Genetic alterations have been frequently found in ovarian cancer. There is some indirect evidence indicating that mutation of the steroid receptor genes may play a role in the carcinogenesis of ovarian cancer. Human androgen receptor (hAR) gene mutations have been found in up to 50% of hormone-relapsed prostate cancer. The role of hAR mutation and its association with decreased expression in ovarian cancer has never been elucidated. In this study mutations of hAR gene in 38 human ovarian cancer cell lines with different AR expression pattern were studied using SSCP. No mutation of the hAR gene was found. Mutation of hAR gene is an infrequent event and therefore unlikely to be involved in the development of ovarian cancer. The decreased expression of hAR in advanced ovarian tumor is not due to genetic aberration of hAR. Mutation screening of hAR may not provide any information for risk assessment of developing ovarian cancer.

Thymidine kinase (TK) gene therapy of solid tumors: valacyclovir facilitates outpatient treatment.

BACKGROUND: In a Phase I study replication-deficient adenovirus containing the herpes simplex virus (HSV) thymidine kinase (TK) gene (AdV-HSV-TK) was instilled intraperitoneally in patients with recurrent ovarian cancer. Patients were treated with Acyclovir (ACV) or Valacyclovir (VCV) as enzymatic substrates. The purpose of this study was to compare serum levels of ACV and VCV. PATIENTS AND METHODS: The antiherpetic prodrug and Topotecan (1.0 mg/m2 over 30 minutes each day for 5 days) were started 24 hours after vector application. Eight patients received ACV (15 mg/kg i.v. over one hour every 8 hours for 42 doses), two patients were started on ACV for 5 days and then switched to oral VCV (2 g every 8 hours for a total of 42 doses). Blood samples were obtained 20 minutes prior to each drug. RESULTS: Serum levels of ACV and VCV (converted to ACV) were comparable. CONCLUSIONS: Suicide gene therapy with TK is under investigation in a variety of solid tumors. Replacing ACV by VCV will offer a cost-effective alternative and will significantly reduce duration of hospital stay improving quality of life and facilitating an outpatient gene therapy concept.

Molecular aspects of ovarian cancer. Is gene therapy the solution?

Genetic abnormalities of cancer cells are complex and usually nonspecific. Genetic anomalies specific to ovarian cancer have not been reported. This article focuses on what molecular anomalies are known in ovarian cancer and describes the first trials that have used transfer of genes to reestablish a normal cellular function in this disease. Suicide gene therapy has been the prototype of this new therapeutic approach.

Telomere length in human white blood cells remains constant with age and is shorter in breast cancer patients.

BACKGROUND: Telomeres, which are TTAGGG repeats at the end of the eukaryotic chromosome, are essential for complete DNA replication. Telomere length has been reported to decrease in peripheral WBC, unlike the telomerase activity found in these cells. The purpose of this study was to investigate whether telomere length in WBC is indeed age dependent and could serve as a genetic marker in breast or ovarian cancer. METHODS: Five age groups: 20-29; 30-39; 4049; 50-59 and > or = 60 years were examined. The cancer patients were 18 women with ovarian cancer and 18 women with breast cancer. Southern blot analysis of the DNA from peripheral white blood cells (WBC) was performed using 32P-labeled (TTAGGG)3 probe. Blots were scanned in a phosphoimager and analyzed by computer-assisted image analysis. RESULTS: No statistically significant correlation was observed between telomere length and age in either healthy females or cancer patients. However, significantly shorter median telomere length was found in WBC obtained from breast cancer patients as compared to healthy individuals and ovarian cancer patients. CONCLUSIONS: It is concluded that telomere length in WBC is not age dependent, but is significantly shorter in breast cancer patients.

Improvement of gene therapy for ovarian cancer by using acyclovir instead of ganciclovir in adenovirus mediated thymidine kinase gene therapy.

Adenovirus(ADV) mediated thymidine kinase(TK) gene therapy followed by ganciclovir(GCV) administration is widely used in different types of cancer. ACV shares the same mechanism of selective cell killing in ADV/TK positive cells as GCV and can be used at 4.5 times higher doses in patients without significant side effects. An increased dose of TK substrate is associated with improved bystander effect and more efficient cell killing. Toxicity and cell killing efficacy were assessed using a 3-(4,5-dimethylthiazol)-2,5-diphenyl tetrazolium bromide(MTT) based assay in three ovarian cancer cell lines with different proliferation patterns. At the same concentration, equal or higher cell killing efficacy and bystander effect were observed using ACV rather than GCV. 2.5 and 5 times (25 micrograms/ml and 50 micrograms/ml) higher concentrations of ACV always resulted in more effective cell killing than GCV (10 micrograms/ml, P < 0.01). Our data indicate that replacing GCV with ACV in the ADV-TK gene therapy may increase the treatment effect without increasing toxicity.

Human epithelial ovarian cancer xenotransplants into nude mice can be cured by adenovirus-mediated thymidine kinase gene therapy.

BACKGROUND: Adenovirus-mediated suicide gene therapy of ovarian cancer has effective anti-tumor effects in vitro and in vivo. By transduction of ovarian adenocarcinoma with the Herpes Simplex Thymidine Kinase gene and subsequent treatment with the antiviral agent ganciclovir, prolongation of survival has been described in nude mice. So far, however, in animal models of solid tumors no cures have been reported after gene therapy. METHODS: In a prospective randomized experimental design 76 mice with xenotransplanted serous ovarian carcinoma were treated with three different doses of ADV/RSV-TK at three different time points followed by intraperitoneal ganciclovir administration. The experiment was designed to show significance of survival differences upon doubling of the number of survived days at a p-value of 0.05 with a power of 80%. The endpoint of the trial was survival. RESULTS: Treatment response was seen in all treated animals evident by significant prolongation of survival. Treatment response was dependent on the therapeutic viral dose and the tumor burden of the animal at the time of treatment. Two out of eight mice with early disease have now survived ten months without evidence of disease with untreated animals dying after nineteen days. Subcutaneous tumor development at the injection site was the reason of death in the remaining six mice of this group. CONCLUSIONS: Intraperitoneal ADV/RSV-TK suicide gene therapy of epithelial ovarian cancer in combination with ganciclovir administration can cure nude mice with early disease. This treatment modality may lend itself to incorporation into the current treatment concept of human ovarian malignancy. Clinical trials are warranted.

Chicken ovalbumin upstream promoter--transcription factor (COUP-TF) expression in human endometrial cancer cell lines.

COUP-TF is an orphan member of the steroid receptor superfamily. COUP-TF down-regulates hormonal induction by other steroid receptors involved in cell proliferation and differentiation. Previous study has suggested a role in gynecological adenocarcinoma. In the present study we evaluated COUP-TF expression in endometrial cancer. Fourteen permanent endometrial cancer cell lines were established front the primary site of 14 endometrial cancer patients. Immunocytochemistry for COUP-TF-like activity was performed using an affinity selected polyclonal rabbit-derived antibody in an immunoperoxidase staining technique. The staining intensity and cell surface area were quantified by image analysis. By immunostain 2 cell lines were COUP-TF (+), 6 (+ +) and 6 (+ + +). Quantitative differences in staining intensity and cell surface area were not significant in these groups. All cell lines tested were immunocytochemically negative for estrogen and progesterone receptors. COUP-TF is a new factor involved in endometrial cancer cell differentiation and growth, especially in estrogen receptor negative tumors.

Adenovirus-mediated thymidine kinase gene transduction in human epithelial ovarian cancer cell lines followed by exposure to ganciclovir.

In an effort to develop gene therapy for ovarian cancer efficacy and toxicity of adenovirus-mediated transfer of the HSV-TK gene followed by administration of ganciclovir were studied in two human epithelial ovarian cancer cell lines Ov-ca-2774 and Ov-ca-1225. 100% transduction was achieved in both cell lines at MOIs of 7 and 15 as demonstrated by X-Gal staining. No toxicity of virus alone was observed at MOIs up to 30. GCV was not toxic up to 200 micrograms/ml. Cell killing efficacy was shown to be dependent on MOI as well as GCV dose. The "bystander effect" of ADV/RSV-TK was quantified by mixing experiments and found to be dependent on the proportion of ADV/RSV-TK positive cells as well as the GCV dosage. Similar results were observed in both cell lines. ADV/RSV-TK mediated gene therapy may be a promising approach in ovarian cancer.

In vivo gene therapy of ovarian cancer by adenovirus-mediated thymidine kinase gene transduction and ganciclovir administration.

Efficacy and toxicity of adenovirus-mediated transfer of the herpes simplex virus thymidine kinase gene followed by administration of ganciclovir were studied in vivo. A human epithelial ovarian cancer animal model was established in nude mice using the serous ovarian adenocarcinoma cell line Ov-ca-2774. Intraperitoneal (ip) injection of 1 x 10(8) Ov-ca-2774 cells resulted in tumor growth and formation of malignant ascites in all 15 animals. In a prospective randomized experimental design mice were treated 1, 3, or 7 days after ip injection of 1 x 10(8) cells with ip injection of 2 x 10(8), 6.7 x 10(8), or 2 x 10(9) pfu ADV.RSV-TK followed by administration of ganciclovir (10 microgram /ml, ip, bid) for 6 consecutive days. End points were survival and toxicity. Mice treated with GCV or HSV-TK alone died from 14.4 +/- 1.7 to 19.5 +/- 3.5 days after treatment as did untreated controls. No toxicity of ADV.RSV-TK was found up to 2 x 10(9) pfu (2 x 10(11) particles). The mice with the highest tumor burden treated with the lowest viral dose lived significantly longer than controls (P < 0.05). Median survival in all other groups of mice treated with ADV.RSV-TK plus GCV was even longer (P < 0.01). Treatment benefit was dependent on ADV/RSV-TK dose and tumor burden. Adenovirus-mediated thymidine kinase gene therapy is a realistic approach to ovarian cancer treatment that warrants investigation in the clinical setting.

Subcellular localization of accumulated p53 in ovarian cancer cells.

Inactivation of the tumor suppressor gene p53 is frequently associated with ovarian cancer. Accumulation of stabilized p53 protein is a common feature in this tumor type. Underlying mutations in the p53 core region can lead to loss of the normal conformational state or loss of residues necessary for DNA binding and transcriptional regulation. Five HPV-free ovarian cancer cell lines established in our laboratory with and without immunocytochemically detectable p53 expression were selected for the correlation of subcellular localization of aberrant p53 and the type of gene mutation. The expression level regarding staining intensity and proportion of cells accumulating p53 was characterized employing an immunoreactive score. Two cell lines with point missense mutations in the core region showed strong nuclear or nuclear plus cytoplasmic staining. One cell line with exclusive staining of the cytoplasm contained a deletion of the major nuclear localization signal. Among two cell lines without p53 accumulation, one contained a microdeletion resulting in a frame shift, the other carried the wild-type sequence. The MDM2 oncogene was not amplified and its gene product was not overexpressed. In ovarian cancer, inactivated p53 can accumulate in both major cell compartments depending on the type of the underlying mutation.