RASAL2 acts as a tumor suppressor in cervical cancer cells

This study was designed to investigate the roles of RASAL2 in cervical cancer(CC).Methods:Fifty-four CC tissues and 33 adjacent tissues were obtained from CC patients admitted to our hospital between March 2012 and June 2014.Real-time polymerase chain reaction and western blotting were performed to analyze the expression of RASAL2 mRNA and protein in these tissues,CC cell lines,and normal cervical cells.Over-expression and silencing of RASAL2 were induced after transfection,and the migration,invasion,and proliferation of the CC cell lines were examined.Results:RASAL2 mRNA and protein expressions were significantly down-regulated in CC tissues and cell lines than in adjacent tissues and normal cervical cells,respectively.While low RASAL2 expression correlated with advanced stage and metastasis of CC,its over-expression significantly inhibited proliferation and metastasis of CC cells and induced apoptosis.Under in vitro conditions,silencing of RASAL2 expression could significantly increase the proliferation,invasion,and migration of CC cells.Conclusion:RASAL2 functioned as a tumor suppressor in CC,and was down-regulated in CC tissue samples and cell lines.

Effect of cisplatin-based concurrent radiochemotherapy on malignant degree of advanced cervical cancer and expression of proto-oncogene and tumor suppressor genes

Objective:To study the effect of cisplatin-based concurrent radiochemotherapy on the malignant degree of advanced cervical cancer and the expression of proto-oncogene and tumor suppressor genes.Methods: A total of 82 patients with advanced cervical cancer who were treated in our hospital between July 2013 and December 2016 were collected and divided into control group and observation group according to random number table, with 41 cases in each group. The control group of patients received radiotherapy alone, while the observation group of patients received cisplatin-based concurrent radiochemotherapy. Tumor marker levels in serum as well as proto-oncogene and tumor suppressor gene expression in tumor tissue were compared between two groups of patients before and after treatment.Results:Before treatment, differences in tumor marker levels in serum as well as proto-oncogene and tumor suppressor gene expression in tumor tissue were not statistically significant between two groups of patients. After treatment, serum tumor markers SCC, CA50, CA724 and CEA levels of observation group were significantly lower than those of control group;proto-oncogene DEK, c-myc and PIK3CA mRNA expression in tumor tissue were significantly lower than those of control group;tumor suppressor genes p53, SOCS-1, FHIT and PTEN mRNA expression in tumor tissue were significantly higher than those of control group.Conclusions:Cisplatin-based concurrent radiochemotherapy can effectively reduce the tumor malignancy and balance the proto-oncogene / tumor suppressor gene expression in patients with advanced cervical cancer.

HSP70 inhibitor combined with cisplatin suppresses the cervical cancer proliferation in vitro and transplanted tumor growth:An experimental study

Objective:To study the regulating effect of HSP70 inhibitor(PES) combined with cisplatin on cervical cancer proliferation in vitro and transplanted tumor growth.Methods:Cervical cancer Hela cell lines were cultured and divided into control group,cisplatin group,PES group and cisplatin+PES group that were treated with serum-free DMEM,cisplatin with final concentration of 10 μmol/L,PES 20 μmol/L and cisplatin 10 μmol/L combined with PES with 20 μmol/L,respectively;animal models with cervical cancer xenografts were established and divided into control group,cisplatin group,PES group and cisplatin+PES group who received intra-tumor injection of normal saline,10 μmol/L cisplatin,20 μmol/L PES as well as 10 μmol/L cisplatin+20 μmol/L PES,respectively.Cell proliferation activity,transplanted tumor volume and mitochondria apoptosis molecule expression were detected.Results:Cell viability value and Bcl-2 mRNA expression in cells of cisplatin group,PES group and cisplatin+PES group were significantly lower than those of control group while Bax,Caspase-3 and Caspase-9 mRNA expression in cells were significantly higher than those of control group;transplanted tumor volume and the Bcl-2 mRNA expression in transplanted tumor tissue of cisplatin group,PES group and cisplatin+PES group were significantly lower than those of control group while Bax,Caspase-3 and Caspase-9 m RNA expression in transplanted tumor tissue were significantly higher than those of control group.Conclusions:HSP70 inhibitor combined with cislatin can inhibit cervical cancer cell proliferation in vitro and transplanted tumor growth through mitochondrial apoptosis pathway.

Anti-tumor effect of matrine combined with cisplatin on rat models of cervical cancer

Objective:To observe the anti-tumor effect of matrine combined with cisplatin on U14 rat models of cervical cancer.Methods:A total of 80 female Kunming rats were used to establish U14 rat models of cervical cancer and then divided into groups Ⅰ,Ⅱ,Ⅲ and Ⅳ,with 20 rats in each.For Group Ⅰ,the control group,injection of normal saline was given around the tumors.For Group Ⅱ,injection of 2 mg/kg cisplatin was given around the tumors.For Group Ⅲ,injection of 75 mg/kg matrine was given around the tumors while the combined injection of matrine and cisplatin was given for Group Ⅳ with the same doses as Groups Ⅱand Ⅲ.The animals were sacrificed 10 d after the injection and tumors were taken out for the comparisons of tumor weights after injection and calculation of anti-tumor rates,while thymus and spleen were taken for thymus index and spleen index.Blood in eyeball was collected for determination of changes in serum creatinine and urea nitrogen levels.Sections of tumor issue were prepared and morphological changes in tumor tissue cells were observed by using immunohistochemistry technique.Results:After injection,the thymus index and spleen index in Groups Ⅲ and Ⅳ were significantly higher than those in Groups Ⅰ and Ⅱ(P<0.05)while the two indexes in Group Ⅱ were significantly lower than Group Ⅰ(P<0.05).The tumor weights in Groups Ⅱ and Ⅳ were significantly smaller than those in Groups Ⅰ and Ⅲ(P<0.05) with significantly higher anti-tumor rates than Groups Ⅰ and Ⅲ(P<0.05).The serum creatinine and urea nitrogen levels in Groups Ⅲ and Ⅳ were significantly lower than Group Ⅱ(P<0.05) and the two indicators in Group Ⅲ were significantly lower than those in Group Ⅳ(P<0.05).The observation under the histological microscope showed densely arranged tumor cells in Group Ⅰ,growing as a crumby structure and diffuse appearance,with hyperchromatic and large nuclei,and abundant cytoplasm.In the case of Group Ⅱ,it showed less tumor cells,with extensive degenerative necrosis,sparse arrangement and karyopyknosis as well as karyoclasis.For Group Ⅲ,necrosis of tumor cells in different sizes and heterogeneous color in nuclei were observed.For Group Ⅳ,the number of tumor cells was significantly smaller than Groups Ⅰ and Ⅲ and the tumor cells presented an appearance of crumby structure as cancer nests,with more proliferation of connective tissue.Conclusions:The treatment of matrine combined with cisplatin can significantly improve the anti-tumor effect on U14 rats with cervical cancer,which can be a new option for the treatment for cervical cancer.

Serum Spondin-2 expression,tumor invasion,and antitumor immune response in patients with cervical cancer

BACKGROUND Cervical cancer is a gynecological malignancy common in middle-aged and older patients,with a high mortality rate.Spondin-2 is an extracellular matrix protein that involved in innate and acquired immune responses.Herein,we investigated the relationship between serum Spondin-2 expression,tumor invasion and infiltration,and immune response in patients with cervical cancer and provided a theoretical basis for clinical practice.AIM To investigate the relationship between serum Spondin-2 expression and cervical cancer-related indicators.METHODS Overall,147 patients with cervical cancer who were admitted to our institution between January 2019 and August 2019 were assigned to the cervical cancer group,and 92 patients with benign uterine lesions and 86 healthy individuals were assigned to the benign and control groups,respectively.In each group,serum Spondin-2 expression was measured,and the receiver operating characteristic(ROC)curve was determined.Patients with cervical cancer were classified into high or low Spondin-2 groups depending on the Spondin-2 threshold value used for diagnosing cervical cancer.Patient’s clinical data were collected to compare the clinicopathologic characteristics,immune cytokine levels,and prognosis of patients with varying Spondin-2 expression levels.RESULTS The expression level of serum Spondin-2 was significantly higher in the cervical cancer group than in the benign and control groups(P<0.05).According to the ROC curve,the cutoff value of Spondin-2 used in the diagnosis of cervical carcinoma was 25.68±7.11μg/L.The proportion of patients with Federation of Gynecology and Obstetrics stage III,nerve invasion,vascular invasion,and lymph node metastasis was higher in the high Spondin-2 group than in the low Spondin-2 group(P<0.05).Interleukin-5(IL-5)and IL-4 Levels were higher in the high Spondin-2 group than in the low Spondin-2 group.In contrast,IL-2 and tumor necrosis factor-αlevels were lower in the high Spondin-2 group than in the low Spondin-2 group(P<0.05).After 3 years of follow-up,progression-free survival and overall survival were significantly shorter in the high Spondin-2 group than in the low Spondin-2 group(P<0.05).CONCLUSION The expression of serum Spondin-2 is upregulated in patients with cervical carcinoma and is related to tumor invasion and infiltration,antitumor immune response,and prognosis.

MR IMAGING ASSESSMENT OF IRREGULAR SHRINKAGE OF TUMOR MORPHOLOGY AND VOLUME IN CERVICAL CANCER DURING RADIATION THERAPY

To study the clinical significance of the morphological and volume changes in cervical cancer during an ongoing course of radiation therapy (RT) using MR imaging. Methods: Serial MR imaging examinations were performed in 60 advanced cervical cancer patients. MR imaging was obtained at the start of RT, at 20-25 Gy (2-2.5 weeks of RT), at 45-50 Gy (4-5 weeks of RT), and 1-2 month post-RT. Tumor morphology was classified qualitatively as well-defined (round/oval with a well-demarcated smooth margin) vs. lobulated vs. irregular and tumor volume was assessed in each serial MR examination independently by ROI volumetry and diameter volumetry. ROI volumetry was traced on the computer workstation with a trackball in each sagittal T2-weighted image and calculated by the summation of all tumor areas in each slice and multiplication by the slice profile. Diameter volumetry was to measure the largest three orthogonal tumor diameters in each orthogonal measurement plane and calculate as an ellipsoid formula (V=d1 x d2 x d3 x p/6). Serial tumor volume was compared between the two measurement methods. Results: The proportion of lobulated and irregular tumors increased early during RT and declined lately post-RT (68% pre-RT, 80% at 2-2.5 weeks of RT, 72% at 4-5 weeks of RT, 33% post-RT). Accordingly, ROI volumetry and diameter volumetry correlated well pre-RT (r1=0.89) and post-RT (r4=0.80), but poorly during RT (r2 = 0.17 at 2-2.5 weeks of RT, r3 = 0.69 at 4-5 weeks of RT). Conclusions: Cervical cancers regress in a non-uniform fashion during RT and undergo increasingly irregular shrinkage. Measurement with ROI volumetry techniques, which can optimally measure irregular volumes, provides better assessment of radiation response during treatment than diameter volumetry.

Effects of preoperative neoadjuvant chemotherapy on malignant biological characteristics in the lesions of patients with ⅠB-ⅡB cervical cancer

Objective: To investigate the effects of preoperative neoadjuvant chemotherapy on malignant biological characteristics in the lesions of patients with ⅠB-ⅡB cervical cancer. Methods:Patients who were diagnosed with ⅠB-ⅡB cervical cancer and underwent surgical treatment in Navy General Hospital of PLA between February 2015 and May 2017 were selected as the research subjects and randomly divided into the neoadjuvant chemotherapy group who underwent preoperative neoadjuvant chemotherapy and the control group who underwent routine preoperative preparation. After surgical resection, the mRNA expression of tumor suppressor genes, proliferation genes as well as migration and invasion genes in cervical cancer tissue were determined by fluorescence quantitative PCR. Results: After surgical resection, tumor suppressor genes THBS2, PTEN, LAST1 and eIF4E3 mRNA expression in cervical cancer tissue of adjuvant chemotherapy group were significantly higher than those of routine surgery group whereas proliferation genes RUNX2, CyclinD1, ALEX1, ALDH1 and ABCG2 as well as migration and invasion genes CXCL12, CXCR4, MMP9, S100A6 and N-cadherin mRNA expression were significantly lower than those of routine surgery group. Conclusion:Preoperative neoadjuvant chemotherapy can inhibit the proliferation, migration and invasion of cancer cells within the lesions of patients with ⅠB-ⅡB cervical cancer.

Effect of Zhenwu decoction combined with paclitaxel neoadjuvant chemotherapy on the malignant degree of cervical cancer

Objective:To study the effect of Zhenwu decoction combined with paclitaxel neoadjuvant chemotherapy on the malignant degree of cervical cancer.Methods: Patients with cervical cancer who accepted neoadjuvant chemotherapy in Nanchong Central Hospital and Suining Central Hospital between March 2015 and June 2017 were selected and randomly divided into two groups, the observation group received neoadjuvant chemotherapy combined with Zhenwu decoction therapy, and the control group received neoadjuvant chemotherapy. Before chemotherapy and after 2 courses of chemotherapy, the serum was collected to detect the contents of tumor markers;after chemotherapy, the surgically removed cervical cancer lesion was collected to detect the mRNA expression of proliferation genes, tumor suppressor genes and invasion genes.Results: After 2 course of chemotherapy, CA12-5, bFGF and SCC-Ag levels in serum of both groups were lower than those before treatment, and CA12-5, bFGF and SCC-Ag levels in serum as well as Ki-67, TK-1, SOX-2, CyclinD1, S100A6, N-cadherin, Gal-3, CatL and CatD mRNA expression in cervical cancer lesion of observation group were lower than those of control group whereas p53, p16, FHIT, NF2-1 and LATS1 mRNA expression in cervical cancer lesions were higher than those of control group.Conclusion:Zhenwu decoction combined with paclitaxel neoadjuvant chemotherapy can be more effective than neoadjuvant chemotherapy alone to kill cervical cancer cells and reduce the malignancy of cervical cancer.

Assessment of the serum tumor markers and lesion cancer cell proliferation after Nimotuzumab combined with cisplatin and radiotherapy treatment of middle-advanced cervical cancer

Objective:To study the change of serum tumor markers and lesion cancer cell proliferation gene expression after Nimotuzumab combined with cisplatin and radiotherapy treatment of middle-advanced cervical cancer.Methods:A total of 78 patients with middle-advanced cervical cancer who were treated in our hospital between August 2013 and February 2016 were collected and divided into the control group (n=39) who received routine radiotherapy and chemotherapy and the observation group (n=39) who received Nimotuzumab combined with cisplatin and radiotherapy according to the single-blind randomized control method, and both groups were treated for 6 weeks. Before treatment and after 6 weeks of treatment, enzyme-linked immunosorbent assay (ELISA) was used to detect serum tumor marker levels, and fluorescence quantitative PCR method was used to detect the lesion proliferation gene mRNA expression.Results: Before treatment, the differences in serum tumor marker levels and lesion proliferation gene mRNA expression were not statistically significant between two groups of group. After 6 weeks of treatment, serum tumor markers SCC, CA125 and CA19-9 levels of observation group were lower than those of control group, lesion pro-proliferation genes B7-H4, HIF-1α, Sp2 and PCNA mRNA expression were lower than those of control group, and lesion anti-proliferation genes PTEN, FHIT and STC1 mRNA expression were higher than those of control group.Conclusion: Nimotuzumab combined with cisplatin and radiotherapy can effectively reduce the serum tumor marker levels, and also inhibit the lesion cancer cell proliferation in patients with middle-advanced cervical cancer.

Diagnostic value of DWI combined with dynamic contrast-enhanced scan of cervical cancer staging弥散加权成像结合动态增强扫描在宫颈癌分期中的诊断价值

目的探讨磁共振弥散加权成像（DWI）结合动态增强扫描在子宫颈癌分期中的诊断价值。方法回顾性分析2013年11月-2014年8月广州医科大学附属第四医院56例经病理证实的宫颈癌患者的DWI及动态增强扫描结果。测量病灶表观扩散系数（ADC）值判断浸润范围，并与手术病理结果进行比较。结果宫颈癌在DWI图像上表现为高信号，ADC值降低，平均ADC值为（0．92±0．16）×10^-3mm^2／s。动态增强扫描早期病灶明显快速强化，延迟期病灶中心呈低信号，病灶边缘呈稍高信号，与邻近正常宫颈组织信号明显不同，形成对比。DWI结合动态增强扫描对宫颈癌FIGO分期（2009）总准确性为93．75％，高于常规MRI分期总准确性（81．25％），对宫旁浸润评估的准确性为100．00％。结论，DWI结合动态增强扫描磁共振检查能提高宫颈癌侵犯程度评价的准确性，对宫颈癌的诊断、临床分期及制订正确的治疗方案具有重要意义。

Current imaging strategies for the evaluation of uterine cervical cancer

Uterine cervical cancer still remains an important socioeconomic issue because it largely affects women of reproductive age.Prognosis is highly depended on extent of the disease at diagnosis and,therefore,accurate staging is crucial for optimal management.Cervical cancer is clinically staged,according to International Federation of Gynecology and Obstetrics guidelines,but,currently,there is increased use of cross sectional imaging modalities [computed tomography(CT),magnetic resonance imaging(MRI),positron emission tomography-CT(PET-CT)] for the study of important prognostic factors like tumor size,parametrial invasion,endocervical extension,pelvic side wall or adjacent/distal organs involvement and lymph node status.Imaging indications also include cervical cancer follow-up,evaluation of tumor response to treatment and selection of suitable candidates for less radical surgeries like radical trachelectomy for fertility preservation.The preferred imaging method for local cervical cancer evaluation is MRI;CT is equally effective for evaluation of extrauterine spread of the disease.PETCT shows high diagnostic performance for the detection of tumor relapse and metastatic lymph nodes.The aim of this review is to familiarize radiologists with the MRI appearance of cervical carcinoma and to discuss the indications of cross sectional imaging during the course of the disease in patients with cervical carcinoma.

Reversal of the immunosuppressive tumor microenvironment via platinum-based neoadjuvant chemotherapy in cervical cancer

Background Immunotherapy favors patients with tumors;however,only 3-26.3%of patients with cervical cancer benefit from single-agent immune checkpoint inhibitors.Combined immunotherapy and chemotherapy has been explored against tumor;however,the combination remains controversial.This study aimed to investigate the tumor immune microenvironment(TIME)and the effects of platinum-based neoadjuvant chemotherapy(NACT)in cervical cancer to identify the clinical value of combining chemotherapy with immunotherapy.Methods Multiplex immunohistochemistry(IHC)with 11 markers(cluster of differentiation[CD]3,CD8,CD4,CD11c,CD68,forkhead box P3[Foxp3],programmed cell death 1[PD-1],programmed cell death 1 ligand 1[PD-L1],indoleamine 2,3-dioxygenase[IDO],cyclin-dependent kinase inhibitor 2A[p16],and cytokeratin[CK])was performed to evaluate TIME from 108 matched pre-and post-NACT cervical cancer samples.The mechanism of antitumor immunity triggered by NACT was explored using RNA sequencing(RNA-seq)from four paired samples and subsequently verified in 41 samples using IHC.Results The infiltration rate of the CD8^(+)T cells in treatment-naive cervical cancer was 0.73%,and those of Foxp3^(+)regulatory T cells(Tregs)and IDO^(+)cells were 0.87%and 17.15%,respectively.Moreover,immunoreactive T cells,dendritic cells,and macrophages were more in the stromal than the intratumor region.NACT increased dendritic,CD3^(+)T,CD8^(+)T,and CD4^(+)T cells and decreased Tregs.The aforementioned alterations occurred predominantly in the stromal region and were primarily in responders.Non-responders primarily showed decreased Tregs and no increase in CD8^(+)T or dendritic cell infiltration.Furthermore,dendritic cells interacted more closely with CD3^(+)T cells after NACT,an effect primarily observed in responders.RNA-seq data revealed activation of the antigen receptor-mediated signaling pathway and upregulation of major histocompatibility complex(MHC)I and MHC II after chemotherapy,validated using IHC.Conclusions NACT can reduce Tregs,and when tumor cells are effectively killed,antigen presentation is enhanced,subsequently activating antitumor immunity finitely.Our study provides the molecular characteristics and theoretical basis for the simultaneous or sequential combination of platinum-based NACT and immunotherapy for cervical cancer.

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.

Micro RNA-378a-3p Downregulation as a Novel Biomarker with Poor Clinical Outcomes in Cervical Cancer

Objective Cervical cancer(CC)is one of the most common malignant tumors in gynecology.This study aimed to investigate the prognostic significance of serum micro RNA(mi R)-378 a-3 p in CC and the effect of mi R-378 a-3 p on tumor growth.Methods Real-time quantitative polymerase chain reaction analysis was used to measure the expression of mi R-378 a-3 p in serum from patients with CC and healthy control subjects as well as from CC tissues and adjacent normal tissues.The association between serum mi R-378 a-3 p levels and clinicopathological factors was analyzed.The correlation between mi R-378 a-3 p levels and overall survival(OS)of CC patients was determined by Kaplan-Meier analysis.The CC cell proliferation and migration abilities after transfection of mi R-378 a-3 p mimics were detected by Cell Counting Kit-8 and scratch wound healing assays,respectively.Tumor volume and weight in mice treated with mi R-378 a-3 p were measured using a caliper and an electronic balance.Results Mi R-378 a-3 p expression was downregulated in the serum and tissues of CC patients compared to that in healthy control subjects and normal tissues,respectively.Low expression of mi R-378 a-3 p was positively correlated with large tumor size,advanced tumor stage,and lymph node metastasis.The OS of patients with low expression of mi R-378 a-3 p was significantly lower than that of patients with high expression.Overexpression of mi R-378 a-3 p suppressed the proliferation and migration of CC cells.In vivo studies indicated that overexpression of mi R-378 a-3 p was associated with decreased tumor volume and weight in mice.Conclusion Mi R-378 a-3 p downregulation is associated with the development and prognosis of CC,suggesting that it may be a potential biomarker for CC.

Effects of high-intensity focused ultrasound combined with interventional chemoembolization on the advanced cervical cancer lesion growth and cell invasion

Objective:To study the effects of high-intensity focused ultrasound (HIFU) combined with interventional chemoembolization on the advanced cervical cancer lesion growth and cell invasion.Methods:Patients with stage IIB-IVA cervical cancer treated in Suining Hospital of TCM between May 2014 and October 2016 were selected and randomly divided into two groups, HIFU group received HIFU combined with interventional chemoembolization, and the control group accepted interventional chemoembolization. The levels of tumor markers in serum as well as the expression of tumor suppressor genes and invasion genes in tumor lesions were determined before and after treatment.Results: 2 weeks, 3 weeks and 4 weeks after treatment, serum TK-1, SCC and CA125 levels of both groups were lower than those before treatment, serum TK-1, SCC and CA125 levels of HIFU group 2 weeks after treatment were not different from those of control group, and serum TK-1, SCC and CA125 levels of HIFU group 3 weeks and 4 weeks after treatment were significantly lower than those of control group;4 weeks after treatment, AIF, NDRG4, SARI and eIF4E3 mRNA expression in tumor lesions of both groups were higher than those before treatment while FAK, KGFR and MMP9 mRNA expression were lower than those before treatment, and AIF, NDRG4, SARI and eIF4E3 mRNA expression in tumor lesions of HIFU group were higher than those of control group while FAK, KGFR and MMP9 mRNA expression were lower than those of control group.Conclusion: HIFU combined with interventional chemoembolization can be more effective in suppressing the advanced cervical cancer lesion growth and cell invasion than interventional chemoembolization alone.

Sanguinarine-Mediated Sensitization of Cervical Cancer SiHa Cells to TRAIL

Introduction: Cervical cancer is primarily caused by the human papilloma virus (HPV), which transforms normal cervical cells into cancerous cells that are highly resistant to radiation and chemotherapy. Induction of apoptosis in transformed cells is a key strategy in successfully treating HPV-induced cervical cancer. TRAIL (tumor necrosis factor related apoptosis-inducing ligand) has been shown to selectively induce apoptosis in cancer cells by binding to death receptors and activating extrinsic pathways for apoptosis. However, certain cervical cancers—such as the cultured cell line SiHa—are remarkably resistant to TRAIL. In this study, SiHa cells were sensitized to TRAIL by using sanguinarine—derived from the plant Sanguinaria Canadensis—which is known to induce oxidative stress and lead to the upregulation of receptors for TRAIL. Methods: Cultured SiHa cells were exposed to sub-lethal doses of sanguinarine in combination with TRAIL. Cell viability changes as well as the production of reactive oxygen species (ROS) were assessed. The induction of apoptosis was investigated by assays for caspase activation. Flow cytometry was performed to analyze expression of death receptors 4/5. Results: Treatment of SiHa cells with a combination of sanguinarine and TRAIL led to a significant reduction in cell viability. Significant increase in ROS was observed and caspase activation assays confirmed the induction of apoptosis. Conclusions: The observed synergistic effect of sanguinarine and TRAIL on SiHa cells is promising for the treatment of cervical, and possibly other, HPV-induced cancers. Oxidative stress caused by sanguinarine seems to play a central role in this synergy. The precise link between reactive oxygen species and the possible upregulation of death receptors needs further investigation. This knowledge will enable us to devise more effective treatments for those who suffer from this devastating disease.

Evaluation of sentinel lymph nodes in vulvar, endometrial and cervical cancers

Sentinel lymph node （SLN） biopsies are a sensitive tool in evaluating lymph nodes for multiple cancers, and in some diseases they decrease morbidity in both the short- and long-term. SLN detection in gynecologic malignancies has been studied extensively over the past decade. We review the current literature on SLN dissection in vulvar, endometrial and cervical cancers. Large, well-designed trials in each of the three types of cancer have demonstrated high sensitivity and low false-negative rates when SLN biopsy is performed in the correct patients and with an appropriate technical approach. In all of these cases the addition of ultra-staging to conventional pathology yields increased detection of micrometastatic disease. Biopsy of the sentinel nodes is feasible and safe in early vulvar malignancies, with multiple studies describing low recurrence rates in those women who have with negative SLNs. There does not appear to be a survival benefit to lymphadenectomy over SLN biopsy and quality of life is improved in women undergoing SLN biopsy. Optimal treatment strategies for women with positive nodal biopsies, particularly in cases with micrometastatic disease, remain unclear. Multiple large studies investigating the utility of SLN biopsy in endometrial malignancy have found that sentinel nodal status is a reliable predictor of metastases in women with low-risk disease. Prospective studies are ongoing and suggest sentinel nodal detection may soon become widely accepted as an alternative standard of care for select cases of endometrial cancer. In cervical cancer, SLN biopsy is accurate for diagnosing metastatic disease in early stage tumors （≤ 2 cm diameter or stage ≤ IB2） where the risk of metastasis is low. It is unknown if women who undergo SLN biopsy alone will have different survival outcomes than women who undergo complete lymphadenectomy in these cases. In a specific population of women with vulvar cancer, SLN dissection is an effective and safe alternative to complete dissection. It can be offered as an alternative management strategy in these women. In women who do undergo SLN biopsy, it is associated with improved quality of life. Promising evidence supporting the utility of SLN dissection in endometrial and cervical cancer continues to emerge, and it may soon become a reasonable option for select patients. However, continued research and refnement of appropriate patient selection and long-term follow-up are necessary.

Effect of preoperative hyperselective uterine arterial chemoembolization on the infiltrative growth of cancer cells in locally advanced cervical cancer

Objective:To study the effect of preoperative hyperselective uterine arterial chemoembolization on the infiltrative growth of cancer cells in locally advanced cervical cancer.Methods: Patients with locally advanced cervical cancer who underwent surgical resection in Ankang Municipality Maternity and Child Care between February 2015 and October 2017 were selected and randomly divided into the observation group who received preoperative hyperselective uterine arterial chemoembolization and the control group who received routine preoperative preparation. The contents of tumor markers in serum were determined at diagnosis and 1 day before undergoing surgery;the expression levels of tumor suppressor genes and invasion genes were determined after surgical resection.Results: 1 day before undergoing surgery, serum CA125, TSGF, SCC and HE4 levels of observation group were lower than those at diagnosis whereas serum CA125, TSGF, SCC and HE4 levels of control group were not significant different from those at diagnosis;after surgical resection, RASSF2A, FHIT, eIF4E3, RIZ1, DAPK and Syk protein expression in cervical cancer lesions of observation group were significantly higher than those of control group;whereas RbAp48, Vimentin, N-cadherin, Sox2,β-catenin and MMP9 protein expression were significantly lower than those of control group.Conclusion: Preoperative hyperselective uterine arterial chemoembolization can inhibit the infiltrative growth of cancer cells in locally advanced cervical cancer.

Long-term survival outcomes and immune checkpoint inhibitor retreatment in patients with advanced cervical cancer treated with camrelizumab plus apatinib in the phase II CLAP study

Background:Camrelizumab plus apatinib have demonstrated robust antitumor activity and safety in patients with advanced cervical cancer(CLAP study;NCT03816553).We herein present the updated long-term results of the CLAP study and explore potential biomarkers for survival.The outcomes of patients who underwent immune checkpoint inhibitor(ICI)retreatment were also reported.Methods:In this phase II trial,eligible patients received camrelizumab 200 mg intravenously every two weeks and apatinib 250 mg orally once daily in 4-week cycles for up to two years.Treatment was continued until disease progression,unacceptable toxicity,or withdrawal of consent.Results:Between January 21 and August 1,2019,a total of 45 patients were enrolled.Data were analyzed as of July 31,2023,representing>48 months since treatment initiation for all patients.Nine(20.0%)patients completed the 2-year study.The median duration of response(DOR)was 16.6 months,and 45.0%of patients achieved a DOR of≥24 months.The 12-month progression-free survival(PFS)rate was 40.7%(95%confidence interval[CI],25.2-55.6),with an 18-month PFS rate of 37.8%(95%CI,22.7-52.8).The median overall survival(OS)was 20.3 months(95%CI,9.3-36.9),and the 24-month OS rate was 47.8%(95%CI,31.7-62.3).Age>50 years,programmed death-ligand 1(PD-L1)combined positive score(CPS)≥1(versus[vs.]<1),CPS≥10(vs.<1),high tumor mutational burden,and PIK3CA mutations were associated with improved PFS(hazard ratio[HR]<1)and longer OS(HR<1).Eight patients who initially responded in the CLAP trial but later experienced disease progression were retreated with ICIs.Among them,2(25.0%)achieved a partial response,while 5(62.5%)had stable disease.Notably,four patients who received retreatment with ICIs survived for more than 45months.No new safety signals were identified in the present study.Conclusion:Long-term survival follow-up data demonstrated that camrelizumab plus apatinib has robust,sustained,and durable efficacy in patients with advanced cervical cancer who progress after first-line platinum-based chemotherapy.No new safety signals were noted with long-term treatment.

Aberrant DNA methylation in cervical carcinogenesis

Persistent infection wit h high-risk types of human papillomavirus(HPV) is known to cause cervical cancer;however,additional genetic and epigenetic alterations are required for progression from precancerous disease to invasive cancer.DNA methylation is an early and frequent molecular alteration in cervical carcinogenesis.In this review,we summarize DNA methylation within the HPV genome and human genome and identify its clinical implications.Methylation of the HPV long control region(LCR) and L1 gene is common during cervical carcinogenesis and increases with the severity of the cervical neoplasm.The L1 gene of HPV16 and HPV18 is consistently hypermethylated in invasive cervical cancers and can potentially be used as a clinical marker of cancer progression.Moreover,promoters of tumor suppressor genes(TSGs) involved in many cellular pathways are methylated in cervical precursors and invasive cancers.Some are associated with squamous cell carcinomas,and others are associated with adenocarcinomas.Identification of methylated TSGs in Pap smear could be an adjuvant test in cervical cancer screening for triage of women with high-risk HPV,atypical squamous cells of undetermined significance,or low grade squamous intraepithelial lesion(LSIL).However,consistent panels must be validated for this approach to be translated to the clinic.Furthermore,reversion of methylated TSGs using demethylating drugs may be an alternative anticancer treatment,but demethylating drugs without toxic carcinogenic and mutagenic properties must be identified and validated.