An improved human tumor stem cell assay in ovarian cancer

To evaluate the effect of improved growth rates of ovarian cancers in the human tumor stem cell assay and its value in predicting clinical chemotherapy response, we studied 59 assays in 54 patients. A total of 81.6% of solid specimens and 85.7% of ascites specimens were successfully cultured and yielded an overall growth rate of 82.9%. Simultaneous primary and metastatic cultures were concordant for chemosensitivity in 80% (n = 16). The patients were evaluated for previous chemotherapy, residual volume of tumor, histologic type, and grade, and these were not statistically different between clinical responders and nonresponders. In vivo-in vitro correlations were made in 27 patients and yielded a predictive response of 13% and predictive resistance of 86% at 70% colony inhibition and 31% and 71% at 50% colony inhibition. Improved growth rates therefore did not result in better predictive correlations. The reported experience in ovarian cancer is summarized and the current status of the human tumor stem cell assay is reviewed.     

Chemotherapy sensitivity of gynecologic malignancies determined by the human tumor stem cell assay (HTSCA)

HTSCA was used to study the chemotherapy sensitivity of gynecological malignant tumors. 34 specimens were obtained from 32 patients. 11 of the 34 specimens (32%) yielded a sufficient number of colonies for testing. On these, 51 separate drug assays were done, each at the 1/10 peak plasma level. 20 drugs were identified as active among 51 drugs tested. Clinical correlations of sensitivity (S) and resistance (R) were studied in 6 patients treated with combination therapies, including drugs selected on the basis of in vitro sensitivity. (formula; see text) We conclude that HTSCA can aid in identifying active drugs for use in designing treatment protocols. However, because of difficulty in preparing a single cell suspension, and the small proportion of tumors suitable for testing, HTSCA will not be used in routine drug sensitivity testing until these drawbacks are eliminated.     

Adjuvant chemotherapy in radical surgery of cervix cancer

Invasive cervical cancer can be treated by surgery, radiotherapy, and cytostatic chemotherapy. For decades, surgery alone or in combination with radiotherapy has been the treatment of choice. Radiotherapy only was reserved for patients with advanced disease. Antineoplastic agents, especially combinations including cisplatin, achieved good results in patients with advanced disease of after other therapeutic modalities had been exhausted. This led us to use postoperative chemotherapy for high-risk patients with positive pelvic or parametrial nodes or vascular invasion. Radiotherapy had not improved the survival of such patients. A combination of bleomycin, vincristine, mitomycin-C and cisplatin and later a combination of carboplatin and bleomycin was used. The results were compared to those of patients with radical abdominal surgery only (N = 118) or with surgery and postoperative radiotherapy (N = 108). The 32 patients who underwent surgery and chemotherapy had statistically higher incidence of all risk factors. Nonetheless, after 4-year follow-up they had less recurrences and deaths than the other patients.     

Glassy cell carcinoma of the uterine cervix: combination chemotherapy with paclitaxel and carboplatin in recurrent tumor

Combination chemotherapy with paclitaxel and carboplatin every 4 weeks for 3 cycles was administered for recurrent glassy cell carcinoma of the uterine cervix in a 67-year-old Japanese female. The response rate was 56% under computed tomography (partial response). However, the effect was transient even with follow-up radiotherapy, and further cases need to be accumulated to determine a successful treatment modality.     

Sertoli cell inactivation by cytotoxic damage to the human testis after cancer chemotherapy

OBJECTIVE: To assess Sertoli cell involvement in postchemotherapy azoospermia. DESIGN: Case report. SETTING: Teaching hospital. PATIENT(S): A 31-year-old azoospermic man who underwent cancer cytotoxic chemotherapy for non-Hodgkin's lymphoma at 13 years of age. INTERVENTION(S): Testicular biopsy specimens were obtained for sperm recovery in preparation for intracytoplasmic sperm injection. The biopsy specimens were evaluated by quantitative immunohistochemistry for the immature Sertoli cell markers cytokeratin 18 (CK-18) and D2-40. MAIN OUTCOME MEASURE(S): Extent of immature Sertoli cells. RESULT(S): A fraction of Sertoli cells (13%) in the atrophic tubules of this patient reexpressed the intermediate filament protein CK-18, which is normally absent after puberty, but not the D2-40 antigen, an Mr 40,000 a-linked membrane glycoprotein, whose loss of expression at puberty marks an irreversible step in Sertoli cell maturation. Tubules with normal spermatogenic progression lined by Sertoli cells negative for CK-18 were also observed. CONCLUSION(S): A fraction of Sertoli cells of this patient initially progressed to full maturation at puberty and reverted to a dedifferentiated state marked by reexpression of CK-18 as a consequence of chemotherapy. This inactivation of Sertoli cells caused by the cytotoxicity of the chemotherapeutic drugs may have contributed to the spermatogenic impairment and resulting infertility.     

Primary chemotherapy and postoperative adjuvant chemotherapy in the treatment of squamous cell carcinoma of the uterine cervix

Two cases with squamous cell carcinoma of the cervix stage IIB with primary tumor mass about 2 X 2 X 2 cm were treated with primary chemotherapy of cis-platinum, 20 micrograms/m2 body surface intravenously daily for 5 consecutive days. Papanicolaou smear turned negative after two courses of chemotherapy in both cases. One of those was followed by radical hysterectomy with lymphadenectomy which confirmed that there was no evidence of disease by histopathology. The third case with stage IIIB and bulky tumor with involvement of pelvic and left common iliac lymph nodes was treated with simple hysterectomy followed by adjuvant chemotherapy with the same medication described above. Complete responses were obtained in all three cases and they are now doing well in a tumor-free state for 26, 30, and 28 months, respectively.     

Intraarterial infusion chemotherapy as a primary treatment in cancer of uterine cervix limited to the pelvis

Percutaneous intraarterial infusion chemotherapy via the internal iliac arteries was performed as a primary treatment in 14 patients with invasive cancer of the uterine cervix. The drugs used were Mitomycin-C, Bleomycin and Cis-Platinum in cycles of 52 hours which were repeated, in some cases, 21 days apart. Complete responses were observed in two patients. Partial responses were seen in 10 patients and no response in two patients. 5 patients were treated surgically and 8 patients received radiotherapy after the infusion. The effect of chemotherapy was more evident in exophytic tumors with less effect on the parametria. Tumor masses can be reduced and patients can then be treated by surgery or radiotherapy. 6 patients died from sepsis.     

Tumor cell kinetics-directed chemotherapy for advanced squamous carcinoma of the cervix

Cell cycle phase specific chemotherapeutic agents may be less effective treatment for metastatic or recurrent cervical carcinoma because most solid tumors contain very low cell growth fractions. Maximum therapeutic advantage may be accomplished through the use of a multiple drug protocol whose schedule is based on phase specificity of drug action and tumor cell kinetics. Preliminary data indicate that administration of a synchronizing agent, followed by cell kinetics analysis using flow microfluorometry techniques, allows a second agent to be scheduled at a time in the cell cycle when it would be most effective. These observations are based on the clinical response of 11 patients treated with kinetics-directed regimens (derived from direct kinetics measurements) or kinetics-based protocols (derived from empirical or historical data).     

激发型CD40单抗对宫颈癌细胞株SiHa化疗敏感性的影响

目的:研究激发型CD40单抗(5C11)对宫颈癌细胞株SiHa化疗敏感性的影响及其机制。方法:采用流式细胞术检测宫颈癌细胞株SiHa上CD40分子的表达,用MTT法检测5C11联合化疗药物对SiHa细胞的作用,PI染色检测SiHa细胞周期的变化,Annexin V-PI法检测细胞凋亡,RT-PCR方法分析单抗5C11作用SiHa细胞后凋亡基因表达水平变化。结果:宫颈癌细胞株SiHa高表达CD40,5C11和盐酸吉西他滨单独抑制细胞生长作用不明显,但两者联合能显著抑制SiHa生长和促进凋亡,SiHa细胞经5C11作用后出现S期阻滞,抗凋亡基因bcl-XL表达明显下调,促凋亡基因Bax的上调作用不明显。结论:CD40分子通过介导S期阻滞和调节凋亡基因表达水平增加SiHa细胞对盐酸吉西他滨化疗的敏感性。     

Effect of combinations of anticancer drugs with interferons on human lung cancer cell lines evaluated by human tumor clonogenic assay]

Using the double agar layer method of human tumor clonogenic assay, the anticancer effect of different combinations of anticancer drugs and interferons was tested on 3 lung cancer cell lines, PC-13, PC-14, and Calu-1. The anticancer drugs and the concentrations used in this study were cisplatin (1.0 microgram/mL), adriamycin (1.0 microgram/mL), mitomycin C (0.2 microgram/mL), VP-16 (5.0 micrograms/mL) and 5-FU (5.0 micrograms/mL). Three kinds of interferon, alpha, beta and gamma in 5,000 units/mL, were tested in combination or in sequence with other anticancer drugs on lung cancer cell lines. The results demonstrate an enhanced anticancer effect on PC-14 only with sequential or simultaneous combination of VP-16 with alpha, beta and gamma interferons; and on Calu-1, only with sequential use of adriamycin and beta-interferon. Our results indicate that there is no unique way of combining anticancer drugs and interferons which can obtain an enhanced anticancer effect on all lung cancer cell lines. The best combination of interferon and anticancer drugs seems to be influenced by the biological characteristics of the cancer cells.     

Carcinoid tumor within a squamous cell carcinoma of the cervix

A small focus of carcinoid tumor within an otherwise typical, poorly differentiated squamous cell carcinoma of the cervix in a 37-year-old female is described. This focus was argyrophilic, but argentaffin negative. There are several papers dealing with primary carcinoid or hormone-secreting tumors of the cervix and, although some of these have shown small foci of epidermoid differentiation, this is the first example in which a small carcinoid focus occurred within and in continuity with a large squamous cell tumor. This may support the theory that APUDomas of the cervix arise from undifferentiated basal stem cells.     

Combination chemotherapy of docetaxel and carboplatin in advanced or recurrent cervix cancer. A pilot study

OBJECTIVES: This is a pilot study for a future trial to assess the efficacy and safety of combination chemotherapy with docetaxel and carboplatin in advanced or recurrent uterine cervix cancer. METHODS: The patients eligible for this study had histologically confirmed, advanced (stage IB2-IV) or recurrent uterine cervix cancer. Eligible patients had measurable lesions and must have sufficient bone marrow, renal, and liver functions. Docetaxel was administered intravenously (IV) at 60 mg/m2 followed by IV carboplatin administration based on AUC = 6. Chemotherapy was repeated in 1-6 courses depending on the purpose of the therapy. The response was evaluated based on RECIST criteria. The toxicity grade was determined by NCI-CTC version 2. RESULTS: During January 2001 and April 2004, 17 patients were entered in this study. The distribution of stage was IB2, 3; IIB, 8; IIIB, 3; IVB, 1; recurrent, 2. There were 9 squamous cell carcinomas, 6 adenocarcinomas, 1 adenosquamous cell carcinoma, and 1 small cell carcinoma. The overall response rate was 76% (2 CR, 11 PR, and 4 SD). No progression of disease was observed. All 5 adenocarcinoma patients in the neoadjuvant chemotherapy group responded including 1 pathological CR. The incidences of grade 3/4 toxicities were 76% for neutrocytopenia, 12% for thrombocytopenia, and 6% for anemia. No grade 3/4 neurotoxicity was observed. CONCLUSIONS: The combination of docetaxel and carboplatin is an effective and safe treatment for uterine cervix cancer. Further evaluation particularly targeted on cervical adenocarcinoma is warranted.     

Squamous cell cancer of the cervix: prognostic factors related to survival

Seven hundred and fifty-three patients with invasive squamous cell cancer of the cervix treated at the University of Michigan from 1970–1985 are reported. These included stage IA 43, stage IB 345, stage IIA 27, stage IIB 163, stage IIIA 4, stage IIIB 113, stage IVA 32, stage IVB 26. The age ranged from 18 to 92 years with a mean of 49.9 years. Clinical characteristics included: nulliparity 11%, married 93%, obese 41%, hypertensive 37%, diabetes 10%, smoking 50%, bleeding 76%. The cumulative five-year survival for all patients was 67% and this was influenced by the stage of disease: stage IA 98%, stage IB 89%, stage IIA 72%, stage IIB 62%, stage III 37%, stage IVA 14%, stage IVB 4%. Patients with a well-differentiated tumor had an 85% survival rate while those with a poorly differentiated tumor had a 57% survival rate. The probability of metastatic disease to lymph nodes corresponded to the stage of disease; stage I 17%, stage II 55%, stage III 70%, stage IV 81%. When lymph nodes were negative, the survival rate for all patients was 86% while those with positive nodes had a 33% survival rate. Factors which influenced survival in the univariate analysis included stage, node status, tumor grade, age, interval from previous pelvic examination, diabetes. Only stage, node status and tumor grade maintained significance in the multiple proportion hazard analysis.     

Localized delivery of chemotherapy to the cervix for radiosensitization

Objective

Chemoradiation is the mainstay of therapy for advanced cervical cancer, with the most effective treatment regimens involving combinations of radiosensitizing agents. However, administration of radiosensitizing chemotherapeutics concurrently with pelvic radiation is not without side effects. The aim of this study was to examine the utility of localized drug delivery as a means of improving drug targeting of radiosensitizing chemotherapeutics to the cervix while limiting systemic toxicities.

Methods

An initial proof-of-concept study was performed in 14 healthy women following local administration of diazepam utilizing a novel cervical delivery device (CerviPrep™). Uterine vein and peripheral blood samples were collected and diazepam was measured using a GC-MS method. In the follow-up study, gemcitabine was applied to the cervix in 17 women undergoing hysterectomy for various gynecological malignancies. Cervical tissue, uterine vein blood samples, and peripheral plasma were collected, and gemcitabine and its deaminated metabolite 2′,2′-difluorodeoxyuridine (dFdU) were measured using HPLC-UV and LC/MS methods.

Results

Targeted delivery of diazepam to the cervix was consistent with parent drug detectable in the uterine vein of 13 of 14 women. In the second study, pharmacologically relevant concentrations of gemcitabine (0.01-6.6 nmol/g tissue) were detected in the cervical tissue of 11 of 16 available specimens with dFdU measureable in 15 samples (0.04-8.8 nmol/g tissue). Neither gemcitabine nor its metabolites were detected in the peripheral plasma of any subject.

Conclusions

Localized drug delivery to the cervix is possible and may be useful in limiting toxicity associated with intravenous administration of chemotherapeutics for radiosensitization.