TGFBI is involved in the formation of polyploid cancer cells and the response to paclitaxel.

BACKGROUND: Most human solid tumors are aneuploid; at the same time, polyploid cancer cells are found to be resistant to radiotherapy and chemotherapy and have a poor prognosis. The transforming growth factor beta induction (TGFBI) protein plays important roles in the development of tumors, depending on the cancer of origin. METHODS: In this study, we established polyploid clones of breast cancer treated with nocodazole. The drug sensitivity was measured by MTT assay. Western blot analysis was used to detect the expression of TGFBI protein in polyploid clones. The effects of paclitaxel on apoptosis, cell cycle and DNA ploidy were analyzed by flow cytometry. TGFBI protein expression was performed in samples from patients with epithelial ovarian tumors by immunohistochemical staining. RESULTS: We found that compared with the MDA-MB-231 cell line, the expression of TGFBI in the HGF1806 cell line was relatively higher. In addition, compared with its parental cells, TGFBI showed relatively low expression in the polyploid breast cancer cell line T-MDA-MB-231. Compared with the empty vector, under paclitaxel treatment, the over-expression of TGFBI in MDA-MB-231 and T-MDA-MB-231 both showed a higher growth inhibition rate. After nocodazole treatment, the over-expression of TGFBI in MDF-MB-231 cells proved that the expression of tetraploid cells was lower compared to the control. The positive rate of TGFBI expression in ovarian cancer specimens before chemotherapy was 33.3% (5/15), which was higher than the positive rate of TGFBI expression in ovarian cancer specimens matched with relapsed specimens after treatment (0%, 0/15). CONCLUSIONS: TGFBI can increase the sensitivity of paclitaxel in polyploid cancer cells and participate in the formation of polyploidy in MDA-MB-231 induced by nocodazole. This newly recognized role of TGFBI provides further insight into the pathogenesis of polyploid cancer and identifies potential new therapeutic targets.

Tracking Neoantigens by Personalized Circulating Tumor DNA Sequencing during Checkpoint Blockade Immunotherapy in Non-Small Cell Lung Cancer.

The evolutionary dynamics of tumor-associated neoantigens carry information about drug sensitivity and resistance to the immune checkpoint blockade (ICB). However, the spectrum of somatic mutations is highly heterogeneous among patients, making it difficult to track neoantigens by circulating tumor DNA (ctDNA) sequencing using "one size fits all" commercial gene panels. Thus, individually customized panels (ICPs) are needed to track neoantigen evolution comprehensively during ICB treatment. Dominant neoantigens are predicted from whole exome sequencing data for treatment-naïve tumor tissues. Panels targeting predicted neoantigens are used for personalized ctDNA sequencing. Analyzing ten patients with non-small cell lung cancer, ICPs are effective for tracking most predicted dominant neoantigens (80-100%) in serial peripheral blood samples, and to detect substantially more genes (18-30) than the capacity of current commercial gene panels. A more than 50% decrease in ctDNA concentration after eight weeks of ICB administration is associated with favorable progression-free survival. Furthermore, at the individual level, the magnitude of the early ctDNA response is correlated with the subsequent change in tumor burden. The application of ICP-based ctDNA sequencing is expected to improve the understanding of ICB-driven tumor evolution and to provide personalized management strategies that optimize the clinical benefits of immunotherapies.

Role of Bcl-2 on drug resistance in breast cancer polyploidy-induced spindle poisons.

Spindle poisons are chemotherapeutic drugs used in the treatment of malignant tumors; however, numerous patients develop resistance following chemotherapy. The present study aimed to induce polyploidy in breast cancer cells using the spindle poison nocodazole to investigate the mechanism of polyploid-induced tumor resistance. It was revealed that the spindle poison nocodazole induced apoptosis in HCC1806 cells but also induced polyploidy in MDA-MB-231 cells. The drug sensitivities of the polyploid MDA-MB-231 cells to paclitaxel, docetaxel, epirubicin, 5-fluorouracil and oxaliplatin were lower than those of the original tumor cells; however, the polyploid MDA-MB-231 cells were more sensitive to etoposide than the original tumor cells. The expression of F-box and WD repeat domain containing 7 (FBW7) was decreased, while the expression of MCL1 apoptosis regulator BCL2 family member (MCL-1) and Bcl-2 was increased, and caspase-3/9 and Bax were not expressed in MDA-MB-231 cells. The resistance to docetaxel and etoposide was reversed, but the sensitivity of paclitaxel was not changed following Bcl-2 silencing. The formation of polyploidy in tumors may be one of the molecular mechanisms underlying tumor resistance to spindle poisons. Expression of the Bcl-2 family members, for example FBW7 and MCL-1, plays a key role in apoptosis and the cell escape process that forms polyploid cells. However, Bcl-2 silencing has different reversal effects on different anti-tumor drugs, which requires further investigation.

Stereotactic Body Radiotherapy Is Effective in Modifying the Tumor Genome and Tumor Immune Microenvironment in Non-Small Cell Lung Cancer or Lung Metastatic Carcinoma.

Background and Purpose: To directly reveal the change in genome mutation, RNA transcript of tumor cells, and tumor microenvironment (TME) after stereotactic body radiotherapy (SBRT) in paired human lung tumor specimens. Materials and Methods: Paired tumor samples were collected from 10 patients with non-small cell lung cancer (NSCLC) or lung metastatic carcinoma within a week before and after SBRT. DNA and RNA of tumor tissues was extracted from the paired samples. Whole-exome and RNA sequencing assays were performed by next-generation sequencing. Gene mutation, genomic expression, T-cell receptor (TCR) repertoire, and profiling of tumor-infiltrating immune cells were analyzed through bioinformatics analysis in paired tumor samples. CD8+ T-cell infiltration and PD-L1 expressions were detected by immunostaining in tumor tissues. Results: The diversity of TCR repertoire and PD-L1 expression increased significantly in the TME, and the most enriched term of the gene ontology analysis was the immune response gene after receiving SBRT. SBRT induced neo-mutation of genes in tumor cells but did not increase tumor mutation burden in tumor tissues. TME displayed complex immune cell changes and infiltration and expression of immune-regulating factors such as C-X-C motif chemokine (CXCL) 10, CXCL16, interferons (IFNs), and IFN receptors. CD8+ T-cells in tumor tissues did not improve significantly after SBRT while the infiltrating TH1 and TH2 cells decreased remarkably. Conclusion: SBRT improved the TCR repertoire diversity and PD-L1 expression in the TME and induced neo-mutation of genes in tumor cells but did not increase CD8+ T-cell infiltration and IFN expression in the tumor tissue within a week.

Ferulic acid ameliorated placental inflammation and apoptosis in rat with preeclampsia.

Preeclampsia (PE) occurs specifically during pregnancy characterized by new-onset hypertension. The pathogenesis of PE was complicated, and inflammation may be central to the pathogenesis of PE. Ferulic acid (FA) is recognized to prevent cell damage and apoptosis induced by oxidative stress and inflammation. In our study, we used NG-nitro-l-arginine methyl ester (L-NAME)-induced rat model of PE to investigate whether FA improved PE and its possible mechanism. We found that FA significantly reduced blood pressure, urine volume, and urinary protein level in rats with PE. Meanwhile, FA decreased L-NAME induced higher expression of circulating TNF-α、IL-6、IL-1ß and PlGF, it reduced placental TNF-α and NF-κB p65. Furthermore, FA rescued L-NAME induced decreasing expression of IL-4 and IL-10 expression in the circulation and placenta of rats. FA also ameliorated placental apoptosis in L-NAME induced rats by increasing Bcl-2 whereas decreasing Bax expression in placenta. It suggested FA as a potential candidate for the treatment of various disorders including L-NAME induced preeclampsia in rats through decreasing placental inflammation and apoptosis.

Diagnosis of fetal megacystis with chromosomal abnormality by 2D prenatal ultrasound: A case report.

RATIONALE: The pathogenesis of fetal megacystis is divided into obstructive and nonobstructive. Megacystis combined with chromosomal abnormalities is rare and most of the cases are nonobstructive. PATIENT CONCERNS: The fetus showed posterior urethral obstructive megacystis with features of bladder enlargement, "keyhole" feature, and thick bladder wall. DIAGNOSES: Here, we present a case of fetal megacystis diagnosed by ultrasound at pregnancy week 15+2 and with multisystem abnormalities. OUTCOMES: Moreover, the fetus showed edema, umbilical cord cyst, cardiac dysplasia, hook-shaped hand, and strephenopodia. These abnormalities strongly suggested chromosomal abnormalities. The fetus was diagnosed with trisomy 18 by amniocentesis. Posterior urethral obstructive megacystis was confirmed by pathology. LESSONS: In conclusion, this case suggests that in the presence of fetal megacystis and multisystem abnormalities, causes should be investigated and the possibility of chromosomal abnormalities should be considered in the presence of multisystem developmental abnormalities.

Acute Fatty Liver of Pregnancy: Analysis on the Diagnosis and Treatment of 15 Cases.

OBJECTIVE: To describe our experience with the diagnosis and management of acute fatty liver of pregnancy (AFLP). STUDY DESIGN: The medical records of pregnant women with AFLP were reviewed for symptoms, laboratory findings, treatment, and maternal and fetal outcomes during a 10-year period between January 2003 and December 2013. RESULTS: During the study period 15 women had AFLP as their discharge diagnosis. The mean gestational age at onset was 36 ± 1 weeks. Eleven cases were primigravidas, and 2 had multiple gestations. All patients presented with nausea, vomiting, and epigastric distress followed by jaundice in the third trimester of pregnancy. Raised transaminases and serum bilirubin as well as coagulopathy were found in all patients (100%), while hypoglycemia was found in 8 patients (53%). Ultrasound examination showed enhanced echo or fatty liver in 11 patients. Maternal morbidity included hepatic encephalopathy (n = 1), renal failure (n = 3), ascites (n = 6), hypoglycemia (n = 8), postpartum hemorrhage (n = 6), upper gastrointestinal hemorrhage (n = 1), and preeclampsia (n = 1). Emergency cesarean section was performed in 12 cases and supracervical hysterectomy was performed in 1 patient due to postpartum hemorrhage and disseminated intravascular coagulation (DIC); all patients survived. All neonates survived except for 2 fetal death cases before admission. CONCLUSION: Early diagnosis and prompt progressive management, including early termination of pregnancy and comprehensive supportive care, are crucial for improving the prognoses of both mother and newborn.

In vitro fertilization complicated by rupture of tubo-ovarian abscess during pregnancy.

OBJECTIVE: Pelvic abscess during pregnancy is an uncommon complication, but can lead to adverse perinatal outcomes during pregnancy. CASE REPORT: We present a patient who developed rupture of a tubo-ovarian abscess during pregnancy following in vitro fertilization and embryo transfer. Thirty-eight reported cases are reviewed, and transvaginal oocyte retrieval, genital tract infections, endometrioma, and previous pelvic surgery are considered as risk factors for pelvic abscess during pregnancy. CONCLUSION: Prolonging gestational duration when an infection situation is allowed is the principle of treatment.

Clinicopathologic study of vulvar Paget's disease in China.

OBJECTIVE: To assess the clinicopathologic characteristics of patients with vulvar Paget's disease who were admitted in our hospital or of cases reported in a Chinese journal. METHODS: The age, disease course, clinical manifestation, pathologic diagnosis, treatment, and follow-up data of patients with vulvar Paget's disease were reviewed. RESULTS: There were 85 cases of vulvar Paget's disease reviewed. The mean age of patients with vulvar Paget's disease was 64.4 years, and their mean disease course was 52.36 months. The patients' primary clinical manifestations were pruritus. Treatment included simple vulvectomy, wide local excision, partial vulvectomy, radical vulvectomy with/without groin dissection, and radiotherapy. Intraepithelial Paget's disease was the most common pathology type (61.5%) followed by invasive Paget's disease (20.0%). There were 7 patients who had a history of secondary malignancy. The mean follow-up period was 43.6 months. The recurrence rate was as high as 43.5%. No significant relationships between margin status, lymph node involvement, pathology type, and recurrence were found (p > .05). CONCLUSIONS: Vulvar Paget's disease has a high local recurrence risk and a long-term follow-up is required.

Endometrial carcinoma in a 15-year-old obese patient with persistent uterine bleeding.

Endometrial carcinoma is the most common malignancy of the upper female genital tract but is rare in teenagers. Here, we report the case of a 15-year-old, nulliparous, morbidly obese female with complaints of asthenia and menometrorrhagia lasting for six months. On examination, the patient had an enlarged uterus approximately 14 gestational weeks in size, and ultrasound revealed an intrauterine mass and polycystic ovaries. An endometrial biopsy performed during hysteroscopy revealed endometrioid adenocarcinoma, and magnetic resonance imaging showed myometrial invasion. The patient underwent a laparotomy involving total abdominal hysterectomy, right salpingo-oophorectomy, wedge-shape dissection of the left ovary, and pelvic and para-aortic lymphadenectomy. We analyze the pathogenesis of endometrial carcinoma in this case and discuss the risk factors for endometrial carcinoma, especially in young women. Gynecologists should be vigilant for persistent abnormal uterine bleeding and other signs of endometrial carcinoma in young women, especially those who have risk factors for the disease.

Exaggerated placental site leading to postpartum hemorrhage: a case report.

BACKGROUND: Exaggerated placental site (EPS) reaction is defined as exuberant infiltration of the endometrium and myometrium at the implantation site by intermediate trophoblastic cells. It is a relatively rare, benign lesion related to pregnancy. The diagnosis of EPS depends mainly on pathologic findings, and it should be distinguished from placental site trophoblastic tumor, placental site nodule and choriocarcinoma. CASE: We present a case of EPS which led clinically to postpartum hemorrhage. During cesarean delivery uterine atony persisted despite pharmacological and surgical intervention. Finally, supracervical hysterectomy was performed due to severe postpartum hemorrhage. CONCLUSION: When postpartum hemorrhage caused by uterine inertia is unresponsive to conventional management, EPS should be considered. Besides timely recognition and intervention with appropriate maneuvers, hysterectomy should be performed as soon as possible to avoid further obstetrical shock and disseminated intravascular coagulation.

The interaction between RAGE gene polymorphisms and HPV infection in determining the susceptibility of cervical cancer in a Chinese population.

AIM: To explore the possible association between the receptor for advanced glycation end products (RAGE) gene polymorphisms and the risk of cervical cancer. METHOD: We enrolled 488 patients with cervical squamous cell carcinoma and 715 age-matched female healthy subjects as controls. Human Papillomavirus (HPV) infection and four RAGE gene polymorphisms (-429T>C, -374T>A, 1704G>T, and 82G>S) in all subjects were determined. RESULTS: The genotype distributions and allele frequencies of -429T>C, 1704 T>G and -374T>A were not significantly different between cervical cancer patients and controls (all P> 0.05). For 82G>S polymorphisms, the genotype distributions and allele frequencies were significantly different between the two groups. The cervical cancer patients had markedly higher percentage of 82SS carriage than controls. The logistic regression analysis showed that the 82SS genotype was associated with significantly elevated risk for cervical cancer, adjusted odds ratio (OR) was 1.98, (P< 0.001). In addition, the 82SS carriers had significantly lower serum soluble RAGE (sRAGE) levels than 82GS and 82GG. The polymorphisms of -429T>C, -374T>A and 1704T>G did not affect the cervical cancer risk and the serum sRAGE levels. When all the cancer patients were stratified by HPV infection status, the 82GS and 82SS genotype carriers in the HPV infection subgroup had increased risk for cervical cancer versus 82GG (OR=1.68 and 1.74, respectively, both P<0.05). This trend was not observed in the subgroup with no detectable HPV DNA. CONCLUSION: Our results suggest that the RAGE 82G>S polymorphisms, interacting with HPV infection, are implicated in the occurrence of cervical cancer.

[Sensitivity to chemotherapeutic drugs of polyploid tumor cells induced by a spindle poison nocodazole].

OBJECTIVE: To investigate the changes of drug sensitivity of spindle poison-induced polyploid tumor cells to chemotherapeutic agents and its possible mechanism. METHODS: Nocodazole in a dose of 100 ng/ml was used to induce polyploidization in a breast cancer cell line MDA-MB-231 cells. The polyploid cells (T-MDA-MB-231) were sorted by flow cytometry. The morphological changes and proliferation of T-MDA-MB-231 cells were compared with that of MDA-MB-231 cells. The cell growth inhibition was assessed by MTT assay. The cells were treated with paclitaxel, docetaxel, vincristine, epirubicin, 5-Fu, VP16 and oxaliplatin, respectively. Those cells were labeled with annexin V-FITC/PI and analyzed by flow cytometry. Bcl-2 was knocked down in T-MDA-MB-231 cells using SiRNA and their growth inhibition was evaluated by MTT assay to evaluate the reversing effect of Bcl-2-silencing on drug resistance. RESULTS: The polyploid T-MDA-MB-231 cells grew in vitro continuously and maintained constant DNA content. They had a larger cell size, and grew more slowly than MDA-MB-231 cells. The IC(50(s)) of T-MDA-MB-231 cells were significantly higher than that of the MDA-MB-231 cells: paclitaxel: (6.37 ± 0.07) vs. (2.05 ± 0.83) µmol/L; docetaxel: (32.98 ± 1.48) vs. (11.95 ± 0.98) µmol/L; vincristine: (35.28 ± 1.66) vs. (14.58 ± 0.94) µmol/L; oxaliplatin: (19.07 ± 0.45) vs. (9.75 ± 1.05) µmol/L; 5-Fu: (85.49 ± 3.21) vs. (31.35 ± 1.51) µmol/L; and epirubicin: (0.53 ± 0.06) vs. (0.15 ± 0.01) µmol/L, (all P < 0.05). The IC(50(s)) of VP16 in T-MDA-MB-231 cells was (2.85 ± 0.50)µmol/L, significantly lower than the (12.20 ± 1.55) µmol/L in MDA-MB-231 cells (P < 0.05), and that of T-MDA-MB-231 cells after Bcl-2-knocked down by siRNA was (19.59 ± 0.48) µmol/L, significantly higher than the (12.20 ± 1.55) µmol/L in the MDA-MB-231 cells (P < 0.05). The IC(50(s)) of docetaxel of T-MDA-MB-231 cells after Bcl-2-knocked down by siRNA was (21.52 ± 0.68) µmol/L, significantly decreased and lower than that before Bcl-2 silencing (32.98 ± 1.48) µmol/L. CONCLUSIONS: Our results indicate that polyploid tumor cells induced by spindle poison Nocodazole are more resistant to most of chemotherapeutic drugs. Downregulation of Bcl-2 increases the sensitivity of polyploid cells to docetaxel. The high expression of Bcl-2 may be one of the drug resistance mechanisms of polyploid tumor cells. The polyploid tumor cells are relatively sensitive to VP16, suggesting that VP16 might be an effective candidate drug for treatment of chemoresistant polyploid tumors.

Clinical outcomes of fertility-sparing treatments in young patients with epithelial ovarian carcinoma.

OBJECTIVE: To assess the clinical outcomes of fertility-sparing treatments in young patients with epithelial ovarian carcinoma (EOC). METHODS: A retrospective study of young EOC inpatients (≤40 years old) was performed during January 1994 and December 2010 in eight institutions. RESULTS: Data were analyzed from 94 patients treated with fertility-sparing surgery with a median follow-up time of 58.7 months. As histologic grade increased, overall survival (OS) and disease-free survival (DFS) of patients receiving fertility-sparing surgery declined. Neither staging surgery nor laparoscopy of early stage EOC with conservative surgery had a significant effect on OS or DFS. Normal menstruation recommenced after chemotherapy in 89% of the fertility-sparing group. Seventeen pregnancies among twelve patients were achieved by the end of the follow-ups. CONCLUSIONS: Fertility-sparing treatment for patients with EOC Stage I Grade 1 could be cautiously considered for young patients. The surgical procedure and surgical route might not significantly influence the prognosis. Standard chemotherapy is not likely to have an evident impact on ovarian function or fertility in young patients.

OPN gene polymorphisms are associated with susceptibility and clinicopatholigical characteristics of cervical cancer in a Chinese cohort.

AIM: To explore the possible association between Osteopontin (OPN) genetic polymorphisms and cervical cancer risk, which remains undocumented yet. METHOD: We enrolled 300 patients with histologically confirmed cervical squamous cell carcinoma and 774 age-matched healthy, unrelated, cancer-free female healthy subjects as control subjects. Three OPN gene polymorphisms were determined. Reulsts: The genotype distributions and allele frequencies of -156 GG/G and -443 T/C polymorphisms were significantly differed between cervical cancer patients and controls. The cervical cancer cases had markedly higher percentage of -156 GG carriage and significantly lower TT and TC of -443 genotypes than controls. The Logistic regression analysis showed that the -156 GG carriage was associated with significantly elevated OR of 2.492 for cervical cancer while the TT and TC of -443 represented lower risks. This trend was not seen in subjects without human papillomavirus infections. In addition, the -156 GG carriages was significantly associated with poorer clinical conditions, including higher clinical stage, poorer tumor differentiation, higher positive lymph node status and higher chance of parametrical invasion. The -443 T/C and -66 T/G polymorphisms did not show any association with the clinicopathological feature. CONCLUSION: These results suggest that the -156 GG/G and -443T/C polymorphisms might be used as a genetic marker for cervical cancer susceptibility.

Screening genes related to development and injury of the mouse optic nerve by cDNA microarrays.

The aim of this study was to screen genes related to the development and injury of the mouse optic nerve so as to provide possible target genes for gene-engineering therapy of central nervous system (CNS) injury. Gene expression was profiled by cDNA microarrays in the mouse superior colliculus at 8-time points during the development or following injury of the optic nerve; consequently, 1,095 highly expressed genes (ratio > or =2) were identified. Then, these genes were categorized functionally; there were 561 genes (51.19%) with unidentified functions and 534 genes (48.81%) with identified or partially identified functions. After discounting the overlapping genes, 486 genes with identified or partially identified functions were categorized into 17 functional groups. The 17 functional groups were as follows: I transcription regulation, II signal transduction, III protein synthesis, IV materials transporting, V RNA processing, VI metabolism-related genes, VII cell cycle or apoptosis-related genes, VIII extracellular matrix, IX protein folding and degradation, X cytoskeleton, XI histone metabolism, XII nervous system specific functional genes, XIII tumor related genes, XIV DNA replication and repair, XV axon growth and guidance, XVI immune response, and XVII cell adhesion. These genes may play key roles in the development, injury, and repairment of the optic nerve.

Heat shock protein 90 and ErbB2 in the cardiac response to doxorubicin injury.

A major drawback to doxorubicin as a cancer-treating drug is cardiac toxicity. To understand the mechanism of doxorubicin cardiac toxicity and the potent synergic effect seen when doxorubicin is combined with anti-ErbB2 (trastuzumab), we developed an in vivo rat model that exhibits progressive dose-dependent cardiac damage and loss of cardiac function after doxorubicin treatment. The hearts of these animals respond to doxorubicin damage by increasing levels of ErbB2 and the ErbB family ligand, neuregulin 1beta, and by activating the downstream Akt signaling pathway. These increases in ErbB2 protein levels are not due to increased ErbB2 mRNA, however, suggesting post-transcriptional mechanisms for regulating this protein in the heart. Accordingly, levels of heat shock protein 90 (HSP90), a known ErbB2 protein stabilizer and chaperone, are increased by doxorubicin treatment, and coimmunoprecipitation reveals binding of HSP90 to ErbB2. Isolated cardiomyocytes are more susceptible to doxorubicin after treatment with HSP90 inhibitor, 17-(allylamino)-17-demethoxygeldanamycin, suggesting that the HSP90 is protective during doxorubicin treatment. Thus, our results provide one plausible mechanism for the susceptibility of the heart to anti-ErbB2 therapy post-doxorubicin therapy in subclinical and clinical conditions. Additionally, these results suggest that further testing is needed for HSP90 inhibitors under various conditions in the heart.

Increased expression of mitotic checkpoint genes in breast cancer cells with chromosomal instability.

PURPOSE: Most breast cancers have chromosomal instability that seems related to defective mitotic spindle checkpoints. Because the molecular basis of this defect is unknown, we evaluated breast cancer cell lines and tissues for possible defects involving the major mitotic checkpoint genes responsible for maintaining chromosomal stability. EXPERIMENTAL DESIGN: We analyzed sequences and expression levels (RNA and protein) of eight major spindle checkpoint genes (MAD1L1, MAD2L1, MAD2L2, BUB1, BUB1B, BUB3, CDC20, and TTK) in a panel of 12 breast cancer cell lines, most with established genetic instability and defective spindle damage checkpoint response. mRNA levels of these genes were also measured in primary tumor samples, and immunohistochemical staining was used to evaluate BUB1B protein levels in a panel of 270 additional cases of breast cancer. RESULTS: No functionally significant sequence variations were found for any of the eight genes in the breast cancer cell lines with chromosomal instability. More surprisingly, the mRNA and protein levels for these checkpoint genes are significantly higher in the genetically unstable breast cancer cell lines and in high-grade primary breast cancer tissues than in the stable (and checkpoint proficient) MCF-10A and normal mammary epithelial cells, or in normal breast tissues. In fact, overexpression of the BUB1B protein is a marker that recognizes nearly 80% of breast cancers in paraffin-embedded tissues. CONCLUSIONS: Defective mitotic spindle checkpoints in breast cancer are most likely not caused by low expression or mutations of these eight checkpoint genes. High levels of these particular transcripts could represent a cellular compensation for defects in other molecular components of the mitotic spindle damage checkpoint, and increased expression of these genes might be markers of breast cancers with chromosomal instability.

[Antisense oligodeoxynucleotides of human telomerse catalytic sub-unit inhibits telomerase activity and proliferation in SKOV3 and COC1].

OBJECTIVE: To study the effects of antisense oligodeoxynucleotides (ODN) of hEST2 (AODN) on telomerase activity and proliferation in ovarian cancer cell lines SKOV3 and COC1. METHODS: Antisense and sense human telomerse catalytic sub-unit (hEST2) phosphorothioate (SODN) and random ODN were designed, synthesized and transfected into SKOV3 and COC1 cells by lipofectamine. The expression of hEST2 mRNA and telomerase activity in SKOV3 and COC1 were tested by reverse transcription-polymerase chain reaction and telomeric repeat amplification protocol before and after transfection. The proliferation and growth in SKOV3 and COC1 were also investigated by methyl thiazolyl tetrazolium and growth curve before and after transfection. RESULTS: AODN could down-regulate the expression of hEST2 mRNA, inhibit telomerase activity and proliferation of ovarian cell lines. The efficiency depends on dose and period of administration. At 48 h, 30 micromol/L AODN had the highest activity. The expression of hEST2 mRNA were declined 54.6% and 44.6% in SKOV3 and COC1 respectively. And also the inhibition of telomerase activity were 47.9% and 42.7% respectively in the two cell lines. CONCLUSIONS: AODN of hEST2 clearly inhibited the proliferation of ovarian cancer cell lines. hEST2 may thus be a new target of gene therapy in ovarian carcinoma.