Labeling of Anti-SAS1B with Zirconium-89 as a Novel Immune-PET Probe for Uterine Cancer Imaging

Overexpression of Sperm Acrosomal SLLP1 Binding protein (SAS1B) in various cancer types, including uterine cancer cells, was discovered a few years ago, and different monoclonal antibodies (anti-SAS1B) that specifically bind to SAS1B antigens were developed. Labeling of these antibodies with radionuclides can provide an opportunity for imaging and radioimmunotherapy. The objective of this study was to label anti-SAS1B (SB5) with Zirconium-89 (89Zr) for PET imaging and determine its biodistribution. Anti-SAS1B (SB5) antibody was labeled with 89Zr indirectly using the chelator desferrioxamine B (DFO), which is currently a best linker for 89Zr. The antibody, SB5, was first conjugated to DFO with a ratio of 1:5 and then labeled with 250 μCi of 89Zr. Results of PET imaging in mouse-bearing uterine cancer tumor showed a limited uptake. The bio-distribution study matched the PET imaging and confirmed the uptake by the tumor, and the accumulation in bones. In conclusion, labeling of anti-SAS1B could provide an effective way of uterine cancer detection and treatment progression.

Risk of critical limb ischemia in long-term uterine cancer survivors:A population-based study

BACKGROUND The risk of critical limb ischemia(CLI)which causes ischemic pain or ischemic loss in the arteries of the lower extremities in long-term uterine cancer(UC)survivors remains unclear,especially in Asian patients,who are younger at the diagnosis of UC than their Western counterparts.AIM To conduct a nationwide population-based study to assess the risk of CLI in UC long-term survivors.METHODS UC survivors,defined as those who survived for longer than 5 years after the diagnosis,were identified and matched at a 1:4 ratio with normal controls.Stratified Cox models were used to assess the risk of CLI.RESULTS From 2000 to 2005,1889 UC survivors who received surgery alone or surgery combined with radiotherapy(RT)were classified into younger(onset age<50 years,n=894)and older(onset age≥50 years,n=995)groups.While compared with normal controls,the younger patients with diabetes,hypertension,and receiving hormone replacement therapy(HRT)were more likely to develop CLI.In contrast,the risk of CLI was associated with adjuvant RT,obesity,hypertension,and HRT in the older group.Among the UC survivors,those who were diagnosed at an advanced age(>65 years,aHR=2.48,P=0.011),had hypertension(aHR=2.18,P=0.008)or received HRT(aHR=3.52,P=0.020)were at a higher risk of CLI.CONCLUSION In this nationwide study,we found that the risk factors associated with CLI were similar in both cohorts except for adjuvant RT that was negligible in the younger group,but positive in the older group.Among the survivors,hypertension,advanced age,and HRT were more hazardous than RT.Secondary prevention should include CLI as a late complication in UC survivorship programs.

Clinical Significance of CENP-H Expression in Uterine Cervical Cancer

Objective This work aims to investigate the expression pattern and clinicopathologic significance of centromere protein H(CENP-H) in uterine cervical cancer(UCC). Methods The level of CENP-H expression in the paraffin sections of 62 UCC cases was determined by the SP immunohistochemical method,with complete clinicopathologic data in all cases.Statistical analysis was conducted to evaluate the prognostic and diagnostic significance of CENP-H using SPSS13.0 software package. Results Immunohistochemical assay showed strong CENP-H expression in 61.29% (38/62) of the paraffin-embedded cervical cancer tissues.Statistical analysis revealed a strong correlation between the CENP-H expression and the clinical classification(P=0.038) of the cervical carcinoma.The expression increased with rise of the stages.The analysis of Cox proportional hazards regression model suggested that CENP-H expression(P=0.002) and tumor stage(P=0.001) were independent prognostic markers for the survival of UCC patients.The survival analysis showed that the survival rate was significantly lower in patients with high expression of CENP-H than in those with low expression of CENP-H(P=0.001). Conclusions CENP-H is likely to be a valuable marker for carcinogenesis and progression of UCC.It might be used as the important diagnostic and prognostic marker for cervical carcinoma patients,especially for those at early stage.

Effects of antisense oligonucleotides targeting VEGF on radio sensitivity of uterine cervix cancer Hela cells

Objective： To determine the impact of antisense oligonucleotides targeting vascular endothelial growth factor （VEGF） on radiosensitivity of uterine cervix cancer Hela cells. Methods： VEGF antisense oligodeoxynucleotides （ASODN） was transfected into Hela cells by liposome-mediated method. Cells transfected with the oligodeoxynuclecotide and saline were used as control groups. Cells were irradiated by 6 MV X ray at the dose of 0 Gy, 2 Gy, 4 Gy and 6 Gy respectively. The expression of VEGF mRNA was determined by RT-PCR. Apoptosis were evaluated using FCM. Cloning efficiency was determined by colony formation assay. Results： The expression of VEGF mRNAwas inhibited by ASODN （P 〈 0.01） in Hela cells. The inhibited activation which was influenced by radiation resulted in increasing apoptosis （P 〈 0.01） and inhibiting plating efficiency （P 〈 0.01）. Conclusion： The expression of VEGF induced by X irradiation in Hela cells can be blocked by VEGF ASODN. Treatment with VEGF might increase apoptosis in HeLa cells and enhance radiosensitivity.

Analysis of the Curative Effect of Preoperative Intra-Arterial Infusion Chemoembolization on Stage IB2-IIB Uterine Cervix Cancer

OBJECTIVETo investigate the short-term and long-termtherapeutic efficacy of preoperative intra-arterial infusion chemo-embolization on stage IB_2-IIB uterine cervix cancer (UCC).METHODS A total of 143 patients with Stage IB_2-IIB UCCwere divided into a clinical trial group and a control group.Thepatients in the clinical trial group(n=86)were treated with acombined therapy,i.e.,preoperative intra-arterial infusion chemo-embolization,surgical therapy and postoperative radiotherapy,and those in the control group (n=57) were given surgical therapyand post-operative radiotherapy.The adverse effects,changes inlocal lesion and pathological examinations of the cancer,and thestate during the surgery were observed after the intra-arterialinfusion chemo-embolization.The survival rate and recurrencerate between the two groups were compared.RESULTS The total effective rate of the intra-arterial infusionchemo-embolization on Stage IB_2-IIB UCC was 93.02%.Thetreatment could reduce tumor size,bring about retro-conversionsof the clinical stage of the tumors and pathological grade of thecancer cells,and decrease the quantity of intra-operative bloodloss as well as the operating time.It could significantly improvethe 5-year survival rate (P<0.05),and reduce the 2 and 5-yeartumor recurrence rates (P<0.05).Moreover,its side effects werelittle.CONCLUSION Preoperative intra-arterial infusion chemo-embolization can create conditions for radical operation,lower thepostoperative recurrence rate,and improve the prognosis in thepatients with UCC.It is an effective therapy in treating UCC.

Potential Anti-cancer Activity of Furanodiene

Objective： To study the isolated from the essential oil VIVO anti-tumor activities of furanodiene of the rhizome of Curcuma wenyujin （C15H200）, a primary sesquiterpene compound YH Chen et C. Ling（Wen Ezhu）, in vitro and in Methods： In vitro MTT assay was used to further study the effects of time and dosage on anti-proliferation of furanodiene against the sensitive Hela, Hep-2, HL-60, U251 cells, based on the cytotoxic effects of furanodiene on 12 human malignant tumor cell lines with the essential oil of Wen Ezhn as control., and the half-inhibitory concentration （IC50） was observed. In vivo uterine cervix （U14） tumor cell was selected and the conventional assay method of anti-tumor activity was employed. Furanodiene liposome was administered intraperitoneally, and tumor-inhibitory rate, thymus and spleen indexes were observed. Results： The inhibitive effects on cell proliferation were shown in all of the twelve cell lines and the cytotoxic effects of furanodiene against Hela, Hep-2, HL-60, U251 cells were observed after 12 h of administration, the effect could last for at least 48 h in a dose dependent manner, and the IC50 values were 0.6, 1.7, 1.8, 7.0μg/ml, respectively. Furanodiene was also found to show inhibitive effects on the proliferation of uterine cervix （U14） tumor induced in mice. The tumor inhibition rates were 36.09% （40 mg/kg）, 41.55% （60 mg/kg）, 58.29% （80 mg/kg）, respectively. Conclusion： Furanodiene is one of primary anti-cancer active components in the essential oil of Wen Ezhu, and also a very effective agent against uterine cervix cancer, and has protection effect on the immune function.

The Establishment and Characterization of a Continuous Cell Line of Mouse Cervical Carcinoma

OBJECTIVE To establish a murine uterine cervical cancer cell line and to define its biological characters. METHODS Transplanted tumor tissue was used for in vitro primary culture of U14 cervical carcinoma cells. After 20 passages, we examined its morphology, chromosomes, tumorigenicity and produced a growth curve. CK was detected by immunohistochemistry, the cell cycle determined by flow cytometry and the metastatic potential assessed in 615 and C57BL/c mice. We also transfected the cells with the pEGFP-N1 plasmid. RESULTS A newly established murine cell line was passaged 50 times over a period of 10 months. The cells grow as a partially suspended culture, and are immunohistochemically CK（＋）. The cell line is characterized by a hypotetraploid karyotype, a chromosomal number of 64-68 and a doubling time of 21.8 h. Exponential growth occurs by the third and forth day of culture. Cell cycle analysis showed G1 34%, G2 26%, and 40% in the S phase. The tumorigenicity was 100% upon implantation. No mycoplasma contamination was detected. A monoclonal continuous U14-GFP cell strain which was 100% GFP （＋） was also produced. CONCLUSION We successfully established a new murine cervical U14 carcinoma cell line and an U14-GFP monoclonal strain. These cell lines are ideal for combined in vivo and in vitro tumor research.

Analysis of Prognosis and Prognostic Factors of Cervical Adenocarcinoma and Adenosqumous Carcinoma of the Cervix

OBJECTIVE To analyze clinical characteristics and treatment methods of the patients with adenocarcinoma of the uterine cervix （AUC） and adenosquamous carcinoma of the cervix （ASCC）. To compare the survival time of the patients in 2 groups and analyze the prognostic factors. METHODS Clinical data of both 123 patients with AUC and 32 patients with ASCC treated at the Cancer Hospital, Chinese Academy of Medical Science （CAMS） ＆ Peking Union Medical College （PUMC）, were retrospectively analyzed. RESULTS The median age of the AUC patients was 50 years, and that of the ASCC patients was 44, P = 0.019. Poorly-differentiated （grade 3） cases accounted for 59.5% of the total ASCC patients, while only 32.5% of the AUC patients were in grade 3, P = 0.002. In 123 AUC patients, relapse or failure of the treatment occurred in 63 of the patients （51.2%）, and the median relapse time was 6 months （0-59 months）. In 32 ASCC patients, relapse or failure of the treatment occurred in 8 of these patients （51.2%）, with a median relapse time of 4.5 months （0-52 months）. The overall 5-year survival rate of the AUC patients was 49.8%, which was significantly lower than that of the ASCC patients （74.1%）, P = 0.015. The 5-year survival rates of the ASCC patients in Stage Ⅰ-Ⅲ were higher than that of the AUC patients with the same stages. However, statistical significant difference could only be found among the patients in Stage II, P = 0.006. The 5-year survival rates of the ASCC patients with various differential grade were higher than those of the AUC patients with the same differential grade, but statistical significant difference could only be found among the patients in the two groups with moderately differentiation, P = 0.039. It was found by Cox regression analysis that only clinical stage （P 〈 0.001） and histological type （P = 0.046） were the independent prognostic factors. CONCLUSION Clinical stage and histological type were the independent prognostic factors of the AUC and ASCC patients. The prOgnosis of ASCC patients is better than that of the AUC patients.

Crosstalk between cell fate and survival pathways during uterine cervical carcinoma progression: a molecular and clinical perspective

The development of uterine cervical cancer is primarily attributed to infection by high-risk human papillomaviruses(HR-HPVs).E5,E6 and E7,the three early oncoproteins of HR-HPVs,have been implicated in initiation and progression of cervical cancer.The intricate molecular mechanisms that orchestrate aberrant cellular transformations to establish carcinoma of the cervical epithelium following viral infections are poorly understood.Here,we discuss how deregulation of three major cell fate regulatory pathways,Hedgehog,Wnt and Notch,and cell survival strategies involving EGFR signaling and G1/S checkpoint contribute towards cervical cancer development and progression.Further exploration of protein interaction database has revealed several genes that are involved in cervical cancer initiation and progression,and the two crucial"driver"genes,MYC and CTNNB1(β-catenin),have been identified as major players in protein-protein interaction network.GSK3βemerged as the key mediator of crosstalk between Hedgehog,Wnt and Notch signaling pathways.GSK3βregulates cytoplasmic stabilization and nuclear translocation ofβ-catenin,which further impacts the expression of MYC,critical for cell cycle progression.Collectively,our analyses suggest that combinatorial therapeutic targeting of these proteins may be more effective in blocking cervical cancer initiation and progression.