Identifying Comprehensive Genomic Alterations and Potential Neoantigens for Cervical Cancer Immunotherapy in a Cohort of Chinese Squamous Cell Carcinoma of the Cervix

Objective Genomic alterations and potential neoantigens for cervical cancer immunotherapy were identified in a cohort of Chinese patients with cervical squamous cell carcinoma(CSCC).Methods Whole-exome sequencing was used to identify genomic alterations and potential neoantigens for CSCC immunotherapy.RNA Sequencing was performed to analyze neoantigen expression.Results Systematic bioinformatics analysis showed that C>T/G>A transitions/transversions were dominant in CSCCs.Missense mutations were the most frequent types of somatic mutation in the coding sequence regions.Mutational signature analysis detected signature 2,signature 6,and signature 7 in CSCC samples.PIK3CA,FBXW7,and BICRA were identified as potential driver genes,with BICRA as a newly reported gene.Genomic variation profiling identified 4,960 potential neoantigens,of which 114 were listed in two neoantigen-related databases.Conclusion The present findings contribute to our understanding of the genomic characteristics of CSCC and provide a foundation for the development of new biotechnology methods for individualized immunotherapy in CSCC.

Mechanisms of Sophora flavescens in the treatment of cervical squamous cell carcinoma based on comprehensive biological analysis,network pharmacology,and experimental verification

Objective:This study used comprehensive bioinformatics analysis and network pharmacology analysis to investigate the potentially relevant mechanisms of Sophora flavescens against cervical squamous cell carcinoma.Methods:Consistently altered genes involved in cervical squamous cell cancerization were analyzed in the GEO database.The chemical ingredients and target genes of Sophora flavescens were explored using the TCMSP database.We obtained the potential therapeutic targets of Sophora flavescens by intersecting the above genesets and validated them in the GEPIA database.The interaction between Sophora flavescens and target genes was predicted by molecular docking.RT-qPCR was used to verify the changes of target genes in HeLa cells treated with Sophora flavescens.Single-gene GSEA functional analysis were performed to determine the molecular mechanisms.Results:Fifteen genes related to the transformation of cervical squamous cell carcinoma were identified,among which AR and ESR1 were confirmed as targets for kaempferol,wighteone,formononetin,and phaseolinon.These compounds are the active ingredients in Sophora flavescens.Low expressions of AR and ESR1 correlate with a poor prognosis,while Sophora flavescens treatment increases the expression of AR and ESR1 in HeLa.GSEA analysis showed that AR and ESR1 mainly participate in the epithelial-mesenchymal transition in cervical squamous cell carcinoma.Conclusion:Sophora flavescens exert anti-tumor effects by targeting AR and ESR1,which may regulate cancer metastasis.

THE DIFFERENTIATION INDUCING EFFECT OF TANSHINONE AND RETINOIC ACID ON HUMAN CERVICAL CARCINOMA CELL LINE（ME180） IN VITRO

The cervical carcinoma cell line, ME180 cells were treated with tanshinone (Tan) or retinoic acid (RA) in DMSO (final concentration 0.02%, V/V) on 4 successive days. The cells treated with the same concentration of DMSO alone served as control. Morphological studies with light and transmission electron microscopy showed that the cells treated with both Tan and RA became welldifferentiated. The cellular growth and proliferation were suppressed (as revealed by cell counting. [3H]-thymidine uptake and colony-forming assay). The number of nuclear organizer regions(AgNORs) in cells reduced and the distribution type returned nearly to normal type. The tumorigenicity in nude mice was reduced. The cell RNA dot hybridization showed that the expression of c-myc and c-Ha-ras mRNA was inhibited markedly. All above results showed that Tan and RA could reverse some malignant Phenotype and possessed differentiation inducing activity on ME180 cell line. No significant difference was observed between the cells treated with Tan and RA.

Expression of CDC42 in cervical squamous cell carcinoma and its correlation with clinicopathologic characteristics

Objective： The high expression of cell division cycle 42 protein （CDC42） may be involved in the occurrence and progression of several tumors. However, the expression and function of CDC42 in cervical squamous cell carcinoma remains unclear. This study aimed to investigate the expression of CDC42 in cervical squamous cell carcinoma and its correlation with clinicopathologic characteristics. Methods： The expression of CDC42 in 162 cervical squamous cell carcinoma tissue samples and 33 normal cervical tissue samples was investigated by immunohistochemistry. The CDC42 mRNA expression was detected by reverse transcription-polymerase chain reaction （RT-PCR）. Results： The cervical squamous cell carcinoma group showed a significantly higher CDC42 positive rate, compared to the normal cervical tissues （P〈0.05）. Fttrthermore, the tissues of stage Ⅱ-Ⅳ carcinoma patients showed higher CDC42 expression levels compared to stage I patients （P=0.05）. In addition, the expression of CDC42 was not correlated to age of patients, differentiation degree of cancer cells, or lymph node metastasis （P〉0.05）. Furthermore, compare with normal cervical tissues, the CDC42 mRNA expression in cervical cancer had no significant difference. Conclusions： CDC42 was up-regulated at protein level, but not mRNA level, in cervical squamous cell carcinoma. The high expression of CDC42 was correlated to the clinical stage of the patients, indicating that CDC42 might contribute to the progression of cervical squamous cell carcinoma.

Coexistence of cervical extramedullary plasmacytoma and squamous cell carcinoma:A case report

BACKGROUND Extramedullary plasmacytoma(EMP),a variant form of myeloma,is a rare solid plasma cell tumor that originates from the bone marrow hematopoietic tissue and accounts for about 3%of all plasma cell tumors.EMP can affect various tissues and organs,about 90%of which is found in the head and neck.However,EMP in the reproductive organs is rare,and is difficult to be distinguished from other primary or metastatic genital tumors according to clinical symptoms and imaging findings.CASE SUMMARY Herein,we report a case with coexistence of EMP and squamous cell carcinoma in the cervix.The first histopathological report of neoplasms on the surface of the cervix and vagina showed an EMP.Both ultrasound and pelvic enhanced magnetic resonance imaging(MRI)indicated that there was a tumor in the cervix.Thus,another cervical biopsy and pathological examination were performed,which indicated EMP combined with squamous cell carcinoma.Then,the patient underwent extensive total hysterectomy(type C1)+systemic lymph node dissection and received 25 external pelvic irradiations with a dose of 50 Gy following surgery.During 2-year follow-up,no recurrence was reported.CONCLUSION In conclusion,EMP involving the reproductive system is relatively rare.In this case,MRI,B-ultrasound,and cervical canal scraping were used to further determine the diagnosis of EMP combined with squamous cell carcinoma.The patient had improved prognosis after appropriate treatments.

Endu combined with concurrent chemotherapy and radiotherapy for stage IIB-IVA cervical squamous cell carcinoma patients

BACKGROUND In recent years,the incidence of cervical cancer has increased with increasing life pressures and changes in women's social roles,posing a serious threat to women's physical and mental health.AIM To explore the clinical effect of Endo combined with concurrent radiotherapy and chemotherapy in the treatment of advanced cervical squamous cell carcinoma.METHODS A total of 120 patients admitted to the oncology department of our hospital were selected as the research subjects.They were equally divided into the test group and the control group(60 patients each)with a random number table.The test group was treated with Endo combined with concurrent radiotherapy and chemotherapy,and the control group was treated with concurrent radiotherapy and chemotherapy.We compared the serum thymidine kinase 1(TK1),human epididymis protein 4(HE4),vascular endothelial growth factor(VEGF),and squamous cell carcinoma-associated antigen(SCC-Ag)levels,the clinical effects and survival before and after radiotherapy and chemotherapy,the quality score,and the 3-year follow-up outcomes between the two groups.RESULTS After chemotherapy,the complete remission+partial remission rate was 85.00%in the test group and 68.33%in the control group;the difference was not statistically significant(P>0.05).Before chemotherapy,the serum TK1,HE4,VEGF,and SCC-Ag levels of the two groups were not significantly different(P>0.05).After chemotherapy,the levels of serum TK1(1.27±0.40 pmol/L),HE4(81.4±24.0 pmol/L),VEGF(235.1±38.0 pg/mL),and SCC-Ag(1.76±0.55 ng/mL)were lower than those in the control group[TK1(1.58±0.51 pmol/L),HE4(98.0±28.6)pmol/L,VEGF(284.2±54.1 pg/mL),and SCC-Ag(2.34±0.78 ng/mL)].The difference was statistically significant(P<0.05).Before chemotherapy,there were no significant differences in the physical,role,mood,cognition,social and symptom scale scores of the two groups(P>0.05).After chemotherapy,the physical,role,mood,cognitive and social scores were higher in the test group than in the control group,and the difference was statistically significant(P<0.05).The symptom scale scores of the test group were all lower than those of the control group,and the difference was statistically significant(P<0.05).The 3-year progression-free survival(PFS)rate was 43.33%in the test group and 26.67%in the control group;the overall survival(OS)rate was 48.33%in the test group and 33.33%in the control group;the differences were not statistically significant(P>0.05).The 3-year PFS time of the test group was 20.0 mo,which was longer than that of the control group(15.0 mo),and the difference was significant(P<0.05).The OS time of the test group was 30.0 mo,which was longer than that of the control group(18.0 mo),and the difference was significant(P<0.05).CONCLUSION Endo combined with concurrent radiotherapy and chemotherapy for the treatment of advanced cervical squamous cell carcinoma has a positive effect on reducing the level of tumor markers in patients,prolonging the PFS and OS times of patients,and improving the quality of life.

THE OVEREXPRESSION AND SIGNIFICANCE OF CYCLIN D1 AND P53 IN CERVICAL SQUAMOUS CELL CARCINOMAS

Objective To investigate the significance of overexpresson of cyclin D1 and P53 protein in cervical squamous cell carcinomas.Methods Fifty cases of invasive cervical squamous cell carcinomas and 10 cases of normal cervical squamous epithelia were investigated with immunihistochemical technique. Results The overexpression of cyclin D1 and P53 in invasive cervical carcinomas was 70% and 50%, respectively. There was no overexpression of them in the control group. The overexpression of cyclin D1 in grade Ⅱ and Ⅲ was much higher than that in gradeⅠ(P<0.05). The overexpresson of cyclin D1 in stage Ⅲ of cervical carcinoma was significantly higher than that in stage Ⅱ (P<0.05). The overexpression of P53 in grade Ⅱ and grade Ⅲ of cervical carcinoma was remarkably higher than that in grade Ⅰ (P<0.05).Conclusion The action point of both cyclin D1 and P53 may be at G1/S transition. The overexpression of them was associated with development and progression of cervical carcinoma probably in different mechanisms and different pathways.

ASSAY OF S100+ LANGERHANS' CELLS IN CERVICAL CARCINOMA

Langerhans cells (LCs) in the normal cervix (control, 19 cases), cervical carcinoma in situ( CIS, 19 cases) and invasive cervical carcinoma (30 cases), interdigitatlng cells (IDCs) and follicular dendritic cells (FDCs) in obturator lymph node draining Invasive cervical carcinoma were quantitatively assessed by ABC immunohistochemical staining using anti-S100 protein antibody. The results indicated that S100+ LCs in sltu, S100+ IDCs and S100+ FDCs In obturator lymph node showed dendritic features with a specific distribution. Number of LCs in situ in Invasive carcinoma increased significantly when compared with CIS and control groups. There was no change in LCs number between grade Ⅱ and Ⅲ of squemous cell carcinoma. Number of IDCs was significantly less in stage Ⅱ than in stage Ⅰ . FDCs number In different Invasive depthes and clinical stages showed no obvious change. The results suggest that progression of cervical carcinoma is closely related to decrease of LCs hi situ and IDCs in regional lymph node. Predominant Immune response of the host to cervical carcinoma should be cellular immunity.

The Number of Positive Pelvic Lymph Nodes and Multiple Groups of Pelvic Lymph Node Metastasis Influence Prognosis in Stage ⅠA-ⅡB Cervical Squamous Cell Carcinoma

Background：Pelvic lymph node metastasis （LNM） is an important prognostic factor in cervical cancer.Cervical squamous cell carcinoma accounts for approximately 75-80% of all cervical cancers.Analyses of the effects of the number of positive lymph nodes （LNs）,unilateral vcrsus bilateral pelvic LNM and a single group versus multiple groups of pelvic LNM on survival and recurrence of cervical squamous cell carcinoma are still lacking.The study aimed to analyze the effects of the number of positive pelvic LNs and a single group versus multiple groups of pelvic LNM on survival and recurrence.Methods：We performed a retrospective review of 296 patients diagnosed with Stage ⅠA-ⅡB cervical squamous cell carcinoma who received extensive/sub-extensive hysterectomy with pelvic lymphadenectomy/pelvic LN sampling at Peking University People＇s Hospital from November 2004 to July 2013.Ten clinicopathological variables were evaluated as risk factors for pelvic LNM：Age at diagnosis,gravidity,clinical stage,histological grade,tumor diameter,lymph-vascular space involvement （LVSI）,depth of cervical stromal invasion,uterine invasion,parametrial invasion,and neoadjuvant chemotherapy.Results：The incidence of pelvic LNM was 20.27% （60/296 cases）.Pelvic LNM （P =0.00） was significantly correlated with recurrence.Pelvic LNM （P =0.00）,the number of positive pelvic LNs （P =0.04） and a single group versus multiple groups of pelvic LNM （P =0.03）had a significant influence on survival.Multivariate analysis revealed that LVSI （P =0.00）,depth of cervical stromal invasion （P =0.00）and parametrial invasion （P =0.03） were independently associated with pelvic LNM.Conclusions：Patients with pelvic LNM had a higher recurrence rate and poor survival outcomes.Furthermore,more than 2 positive pelvic LNs and multiple groups of pelvic LNM appeared to identify patients with worse survival outcomes in node-positive ⅠA-ⅡB cervical squamous cell carcinoma.LVSI,parametrial invasion,and depth of cervical stromal invasion were identified as independent clinicopathological risk factors for pelvic LNM.

A preliminary study of genes related to concomitant chemoradiotherapy resistance in advanced uterine cervical squamous cell carcinoma

Background Tumor intrinsic chemoradiotherapy resistance is the primary factor in concomitant chemoradiotherapy failure in advanced uterine cervical squamous cell carcinoma. This study aims to identify a set of genes and molecular pathways related to this condition.

Polarimetry feature parameter deriving from Mueller matrix imaging and auto-diagnostic signicance to distinguish HSIL and CSCC

High-grade squamous intraepithelial lesion(HSIL)is regarded as a serious precancerous state of cervix,and it is easy to progress into cervical invasive carcinoma which highlights the importance of earlier diagnosis and treatment of cervical lesions.Pathologists examine the biopsied cervical epithelial tissue through a microscope.The pathological examination will take a long time and sometimes results in high inter-and intra-observer variability in outcomes.Polarization imaging techniques have broad application prospects for biomedical diagnosis such as breast,liver,colon,thyroid and so on.In our team,we have derived polarimetry feature parameters(PFPs)to characterize microstructural features in histological sections of breast tissues,and the accuracy for PFPs ranges from 0.82 to 0.91.Therefore,the aim of this paper is to distinguish automatically microstructural features between HSIL and cervical squamous cell carcinoma(CSCC)by means of polarization imaging techniques,and try to provide quantitative reference index for patho-logical diagnosis which can alleviate the workload of pathologists.Polarization images of the H&E stained histological slices were obtained by Mueller matrix microscope.The typical path-ological structure area was labeled by two experienced pathologists.Calculate the polarimetry basis parameter(PBP)statistics for this region.The PBP statistics(stat PBPs)are screened by mutual information(MI)method.The training method is based on a linear discriminant analysis(LDA)classier whichnds the most simplied linear combination from these stat PBPs and the accuracy remains constant to characterize the specic microstructural feature quantitatively in cervical squamous epithelium.We present results from 37 clinical patients with analysis regions of cervical squamous epithelium.The accuracy of PFP for recognizing HSIL and CSCC was 83.8%and 87.5%,respectively.This work demonstrates the ability of PFP to quantitatively charac-terize the cervical squamous epithelial lesions in the H&E pathological sections.Signicance:Polarization detection technology provides an effcient method for digital pathological diagnosis and points out a new way for automatic screening of pathological sections.