Knockdown of circular RNA (CircRNA)_001896 inhibits cervical cancer proliferation and stemness in vivo and in vitro

Objective:Previous studies indicated that aberrant circular RNA(circRNA)expression affects gene expression regulatory networks,leading to the aberrant activation of tumor pathways and promoting tumor cell growth.However,the expression,clinical significance,and effects on cell propagation,invasion,and dissemination of circRNA_001896 in cervical cancer(CC)tissues remain unclear.Methods:The Gene Expression Omnibus(GEO)datasets(GSE113696 and GSE102686)were used to examine differential circRNA expression in CC and adjacent tissues.The expression of circRNA_001896 was detected in 72 CC patients usingfluorescence quantitative PCR.Correlation analysis with clinical pathological features was performed through COX multivariate and univariate analysis.The effect of circRNA_001896 downregulation on CC cell propagation was examined using the cell counting kit-8(CCK-8)test,clonogenic,3D sphere formation,and in vivo tumorigenesis assays.Results:Intersection of the GSE113696 and GSE102686 datasets revealed an increased expression of four circRNAs,including circRNA_001896,in CC tissues.Fluorescence quantitative PCR confirmed circRNA_001896 as a circular RNA.High expression of circRNA_001896 was considerably associated with lymph node metastasis,International Federation of Gynecologists and Obstetricians(FIGO)stage,tumor diameter,and survival period in CC patients.Proportional hazards model(COX)univariate and multivariate analyses revealed that circRNA_001896 expressions are a distinct risk factor affecting CC patients’prognosis.Cellular functional experiments showed that downregulating circRNA_001896 substantially suppressed CC cell growth,colony formation,and 3D sphere-forming ability.In vivo,tumorigenesis analysis in nude mice demonstrated that downregulating circRNA_001896 remarkably reduced the in vivo proliferation capacity of CC cells.Conclusion:CircRNA_001896 is highly expressed in CC tissues and is substantially related to lymph node metastasis,FIGO stage,tumor size,and survival period in patients.Moreover,downregulating circRNA_001896 significantly inhibits both in vivo and in vitro propagation of CC cells.Therefore,circRNA_001896 might be used as a biomarker for targeted therapy in cervical cancer.

Subgroups of peripheral immune effector cells in cervical cancer patients are more sensitive to radiation therapy than chemotherapy

Background:CD8 positive T lymphocytes and natural killer(NK)cells in the peripheral blood of cervical cancer patients exhibit varying sensitivities to radiotherapy and chemotherapy.Methods:A total of 50 healthy peoples and 60 cervical cancer patients were recruited.The patients with cervical cancer were separated into two groups:radiation and chemotherapy,and blood sample were collected before and after treatment.Data on the proportion of CD8 positive T lymphocytes and NK cells were gathered for analytical evaluation.Results:Compared to healthy individuals,patients with cervical cancer exhibit a reduced proportion of CD8 positive T cells within their peripheral blood.And for patients with cervical cancer,radiation therapy has been found to be more effective than chemotherapy in increasing the proportion of CD8 positive T lymphocytes and NK cells.Conclusions:These results suggest that radiation therapy increases the levels of CD8 positive T lymphocytes and NK cells within the peripheral blood of patients with cervical cancer.The study hypothesis that the changes in the percentage of CD8 positive T lymphocytes may serve as a potential indicator for predicting treatment efficacy.

Relationship between Circulating Plasma Galectin-3 Levels and T-Cell Activation during Cervical Cancer Chemotherapy

Objective: Despite the existence of several therapeutic strategies, the management of cervical cancer remains challenging. Our region has very little data on the interaction between the immune system and the clinical response to chemotherapy. This work examines plasma levels of galectin-3 (Gal-3) and percentages of activated T cells in patients with cervical cancer treated with chemotherapy and investigates if there is a relationship between the rates of these two elements. Methods: We compared data from 37 patients with cervical cancer undergoing chemotherapy and 42 controls with normal cervical cytology. Plasma Gal-3 concentrations were assessed by ELISA and expression of activation markers by T cells (CD69 and HLA-DR) was assessed by flow cytometry at three different time points during chemotherapy. Results: Our results showed that patients had a significantly higher concentration of Gal-3 compared to controls (4.025 vs. 1.340, p 0.001), similarly, they had a significantly high percentage of activated lymphocytes (2.610 vs. 0.731;p 0.0001). According to the response to treatment, patients with no response to treatment had a lower concentration of circulating Gal-3 but had approximately the same percentage of activated CD4 and CD8 lymphocytes as patients with a partial or total response. In addition, we found a positive correlation between the Gal-3 level and CD4 T cells expressing the activation marker CD69 (p 0.05;rho = 0.44). Conclusion: In conclusion, our results show that there would be a relationship between circulating galectin-3 and the percentage of peripheral CD4+CD69+ cells in cervical cancer.

Quantitative Detection of Screening for Cervical Lesions with ThinPrep Cytology Test

OBJECTIVE To investigate the available parameters in gynecological screening for cervical lesions by liquid-based cytology technology （ThinPrep Cytology Test, TCT） and The Bethesda System （TBS）, also with computer image analysis. METHODS With application of the image analysis system, all grades of cervical lesion cells were detected quantitatively and sorted in atypical squamous cells of undetermined significance （ASCUS）, atypical squamous cells-cannot exclude HSIL （ASC-H）, low-grade squamous intraepithelial lesion （LSIL）, high-grade squamous intraepithelial lesion （HSIL） and cervical squamous cell carcinoma （SCC） with the mean optical density （MOD）, average grey （AG）, positive units （PU）, and nucleus to cytoplasmic ratio （N： C）. Differences between each group of cells were compared and analyzed statistically. RESULTS Apart from four stereologic parameters in LSIL and HSIL groups there were no differences among them, in the other groups, there was statistically significant in differences between MOD, AG and PU values. Differences between them in the ratio of nucleus to cytoplasm were highly statistically significant. CONCLUSION Stereological indexes may serve as a screening tool for cervical lesions. The image analysis system is expected to become a new means of cytological assisted diagnosis.

The delay, symptoms, and survival of Ivorian adolescent girls and young adults with uterine cervical cancer

Objective:To generate data on the nature and duration of cervical cancer symptoms,risk factors for delayed consultation,and diagnosis of adolescent and young adult groups in Cote d'Ivoire.Methods:This is a hospital-based cross-sectional study,conducted from July 2012 to May 2018,at the Department of Gynecology of the Yopougon teaching hospital in Cote d’Ivoire.The inclusion criteria were those who were under 40 years of age at the time of diagnosis with oral and written consent.Those whose information was more than 10%insufficient for the standards were excluded from the study.The survey files standardized anonymous pre-established data collected using medical records followed by phone calls.Results:The average age of the participants was 34±4.95 years with a minimum of 21 years.The mean parity per woman was 3.49±3.54.34.9%were HIV positive.The median patient delay was 122 days with a long delay in 84.62%of patients.The median time to total diagnosis was 209 days with a longer total delay in diagnosis of 87.18%.41%of patients honored their treatment,68.75%of which had received surgery,and 31.25%of which had received chemotherapy.The 5-year survival rate was 65%.Conclusion:The Ivorian health system must focus on primary and secondary prevention,the only guarantee to decrease the morbidity and mortality indicators linked to cervical cancer.

Prevalence of high-risk human papillomavirus infection in different cervical lesion among organized health-examination women in Shanghai, China

Background Cervical cancer is the second most common cause of death from cancer among women worldwide. Human papillomavirus （HPV） plays a central role in the etiology of cervical cancer. It is important to describe the prevalence of HPV infection in different types of cervical lesions and to explore the relation between HPV viral load and the severity of cervical lesions. Methods To describe the HPV infection prevalence and viral load in different age groups, we retrospectively investigated 6405 cases of women who were organized by their units to take health-examination. They were given Hybrid Capture II tests between January 2005 and December 2006. The correlation between HPV viral load and pathology was assessed. Results Overall HPV infection prevalence was 29.1% （1864/6405）, while in women 18-20 years old it was 54.4% （31/57）, the highest among all age groups. After declining rapidly, HPV prevalence stabilized at about 30.0% in women aged 30 and older. Of the 6405 women, 1483 women had a colposcopic biopsy and 33.2% （492/1483） were positive for HPV DNA. Twenty-one percent of women with a normal diagnosis （238/1095） had HPV infection, a statistically significantly lower prevalence than in women with cervical lesions, including those with cervical intraepithelial neoplasia （68.8% in CIN1,66.7% in CIN2, and 76.5% in CIN3） or with cervical cancer （94.1%）. The correlation coefficient between viral load and cervical lesion severity was 0.134, which was not statistically significant （P=0.075）. Viral load values in women with CINs and cervical cancer were calculated, and no significant differences were identified. Conclusions The prevalence of high-risk HPV infection among women attending hospitals for health-examination in Shanghai is similar to the worldwide rate. HPV viral load can distinguish cervical lesions from normal individuals but cannot adequately predict the severity of cervical lesions.

Overexpression of Bcl-2 partly inhibits apoptosis of human cervical cancer SiHa cells induced by arsenic trioxide

OBJECTIVE: To study the biological effect of arsenic trioxide (As2O3) on human cervical cancer SiHa cells and SiHa cells overexpressing bcl-2 gene. METHODS: SiHa cells with overexpression of Bcl-2 (SiHa-Bcl2 cells) were established by transfecting SiHa cells with Bcl-2 expression vector. The sensitivities of SiHa and SiHa-Bcl2 cells to As2O3 were determined using MTT (Thiazolyl blue) reduction and colony forming ability assay, morphological analysis, flow cytometric analysis, DNA agarose gel electrophoresis, in situ cell death detection (TUNEL), Northern blot, RT-PCR and Western blot. RESULTS: As2O3 inhibited the growth of SiHa cells and induced G2/M arrest and apoptosis of the cells. RT-PCR and Western blot analysis revealed that As2O3 induced SiHa cell apoptosis possibly via inhibiting the expression of HPV16 E7 and decreasing the expression of c-myc. However, we found that SiHa-Bcl2 cells partly resisted As2O3 induced apoptosis, which might be related to the prevention of the down-regulation of HPV16 E7 and c-myc gene expression. Nevertheless, As2O3 at a high concentration could still induce apoptosis of SiHa-Bcl2 cells mainly via decreasing Bcl-2 expression and slightly inhibiting viral gene expression. CONCLUSION: As2O3 is an inducer of the apoptosis of human cervical carcinoma cells and the cells overexpressing Bcl-2 can partly resist As2O3 induced apoptosis, but the exact mechanism is unclear.

Therapeutic observation of acupuncture plus navel acupuncture for urinary retention after radical hysterectomy for cervical cancer

Objective: To observe the clinical efficacy of acupuncture plus navel acupuncture for patients with urinary retention after radical hysterectomy for cervical cancer. Methods: A total of 64 patients with urinary retention after radical hysterectomy for cervical cancer was divided into a navel acupuncture group (22 cases), an acupuncture group (18 cases) and an acupuncture plus navel acupuncture group (24 cases). All three groups received bladder function training and neuromuscular electrical stimulation. In addition, navel points were combined in the navel acupuncture group. Electroacupuncture was conducted to Qihai (CV 6), Zhongji (CV 3), Dahe (KI 12), Shuidao (ST 28), Ciliao (BL 32) and Huiyang (BL 35) in the acupuncture group. The acupuncture plus navel acupuncture group received both treatments. The catheter was removed after 3 d of treatment. Spontaneous urination, residual urine volume, urinary catheter dependence and recurrence after 3 d, 6 d and 9 d of treatment in each group were observed, respectively. Results: In the acupuncture plus navel acupuncture group, the markedly effective rates after 3 d, 6 d and 9 d of treatment were significantly higher than those in the navel acupuncture group and the acupuncture group;the urinary catheter dependence was lower than that of the other two groups, and the differences were statistically significant (P<0.05, P<0.01);the spontaneous urination time was shorter than that of the navel acupuncture group and the acupuncture group, and the differences were statistically significant (P<0.05, P<0.01);the residual urine volume was significantly less than that of the navel acupuncture group and the acupuncture group, and the differences were statistically significant (both P<0.01). After the catheter was removed, recurrence was observed from the next day after spontaneous urination was resumed. There were 2 cases of recurrence in the navel acupuncture group, 2 cases in the acupuncture group and 1 case in the acupuncture plus navel acupuncture group. The recurrence rate of the acupuncture plus navel acupuncture group was significantly lower than that of the navel acupuncture group and the acupuncture group (both P<0.01). Conclusion: Acupuncture plus navel acupuncture has satisfactory efficacy for urinary retention after radical hysterectomy for cervical cancer. It can significantly shorten the urinary retention time, reduce the patient's dependence on urinary catheter, and reduce the residual urine volume.

Efficacy observation on acupuncture-moxibustion for urinary retention after surgery for cervical cancer

Objective：To observe the efficacy of acupuncture-moxibustion therapy in treating urinary retention after surgery for cervical cancer. Methods：Fifty eligible patients were randomized into a treatment group and a control group by their visiting sequence, 25 in each group. The control group was intervened by indwelling urethral catheter after gynecological surgery; while the treatment group started to receive acupuncture-moxibustion and cupping treatment at the 7th day after surgery, in addition to the intervention given to the control group. The residual urine volume and total effective rate of the two groups were detected. Results：After 7-day treatments, the total effective rate was 92.0% in the treatment group versus 72.0% in the control group, and the difference was statistically significant （P〈0.05）. Conclusion：Acupuncture-moxibustion treatment plus cupping based on the conventional intervention is effective for urinary retention after surgery for cervical cancer, worth promoting in clinic.

Effect of acupuncture plus medium-frequency electric stimulation on bladder function after radical hysterectomy for cervical cancer

Objective： To observe the effect of acupuncture plus medium-frequency electric stimulation on bladder function after radical hysterectomy for cervical cancer. Methods： A total of 170 patients confirmed by biopsy and surgical pathology and treated with radical hysterectomy for cervical cancer were randomly divided into an acupuncture group, an observation group and a control group by random digital table. Fifty-six cases in the acupuncture group were treated with acupuncture after surgery, one session each day, till the extubation day; 58 cases in the observation group were treated with acupuncture plus medium-frequency electric stimulation after surgery, one session each day, till the extubation day; and 56 cases in the control group didn＇t receive any intervention, and the catheter was retained till the extubation day. On the postoperative day 14, all groups were dealt with catheterization to determine the residual urine volume, in order to observe the occurrence rate of urinary retention and the recovery of bladder function and to compare the differences of the therapeutic effects among the groups. Results： The occurrence rates of urinary retention were respectively 16.1%, 20.7% and 46.4% in the acupuncture group, observation group and control group, with significant differences （P〈0.05）. When comparing the acupuncture group and the observation group with the control group, there were significant differences （both P〈0.05）. In comparison between the acupuncture group and the observation group, there was no significant difference （P〉0.05）. The recovery rates of bladder function in the three group were respectively 42.9%, 60.3% and 41.1% without significant differences （P〉0.05）. Moreover, there were no significant between-group differences （P〉0.05）, but acupuncture plus medium-frequency electric stimulation showed a better tendency. Conclusion： The single application of acupuncture therapy is effective and able to prevent urinary retention after radical hysterectomy for cervical cancer. Acupuncture plus medium-frequency electric stimulation can effectively enhance the recovery of bladder function and reduce the residual urine volume in the bladder to the maximum.

Effects of Moxibustion on Hemoglobin and Immunoglobulin in Cervical Cancer Patients Undergoing Radiotherapy

Objective： To observe the effects of moxibustion on hemoglobin and immunoglobulin in cervical cancer patients on radiotherapy. Methods： Sixty-six patients with cervical cancer were randomized into a moxibustion group （n=36） and a control group （n=30）, and underwent the same radiotherapy; during the radiotherapy period, the moxibustion group was given moxibustion at Shenque （CV 8） and Sanyinjiao （SP 6）, and the control group was given no other treatment. Agar diffusion method was employed to detect the concentrations of hemoglobin and immunoglobulin in peripheral blood before and after treatment. Results： After eight-week treatment, the concentrations of hemoglobin and immunoglobulin were higher in the moxibustion group than those in the control group, with a significant difference in the concentration of IgG between the two groups （P〈0.01）. Conclusion： Moxibustion can increase the concentrations of hemoglobin and immunoglobulin in cervical cancer patients on radiotherapy to improve the immunity.

Identification of biomarkers for cervical cancer in peripheral blood lymphocytes using oligonucleotide microarrays

Background Oligonucleotide microarrays are increasingly being used to identify gene expression profiles that associated with complex genetic diseases. Peripheral lymphocytes communicate with cells and extracellular matrixes in almost all tissues and organs in human body, suggesting that the gene expression profiles in peripheral lymphocytes may reflect the presence of disease in the body. This study aimed to identify molecular biomarkers for cervical cancer in peripheral blood lymphocytes by using oligonucleotide microarrays. Methods Total RNA was extracted from peripheral blood lymphocytes of 24 early stage cervical cancer patients and 18 healthy controls. We used 22K Human Genome microarrays to profile peripheral blood lymphocytes from 4 early stage cervical cancer patients and compared their gene expression profiles with those from 3 healthy controls. Differentially expressed genes would be identified if they had adjusted P values of less than 0.05 and a groupwise average fold change greater than 1.5 or less than 0.67. Then the selected 5 genes were validated in the remaining 20 early stage cervical cancer patients and the 15 healthy controls by using real-time reverse-transcription polymerase chain reaction （RT-PCR）. Results Genes identified by the gene selection program expressed differently between the blood samples of the early stage cervical cancer patients and those of the healthy controls. To validate the gene expression data, 5 genes were analyzed by real-time RT-PCR. In three of the 5 identified genes, tenasin-c （TNC）, nuceolin （NCL）, and enolase 2 （EN02） showed a significant up-regulation in the blood samples of the early stage cervical cancer patients versus that of the healthy controls. Conclusions The up-regulation of TNC, NCL, and EN02 in peripheral blood may be used to identify novel blood biomarkers for detecting cervical cancer in a clinically accessible surrogate tissue, and thus to provide a possibility to develop a noninvasive and predictive diagnosis for the disease.

Clinical study on intradermal needle therapy in treating urinary retention after cervical cancer surgery

Objective:To observe the clinical efficacy of intradermal needle therapy for urinary retention after cervical cancer surgery.Methods:A total of 100 patients with urinary retention after cervical cancer surgery were randomized into a control group and an observation group,with 50 cases in each group.The control group was treated with basic nursing only,and the observation group was treated with additional intradermal needle therapy.Both groups were treated for 2 courses of treatment.The main symptom scores and residual urine volume of the two groups were observed before and after treatment,and the inpatient time,catheter indwelling time and the clinical efficacy were compared between the two groups.Results:The total effective rate was 96.0%in the observation group and 88.0% in the control group,and the difference between the two groups was statistically significant(P<0.05).After treatment,the main symptom scores and residual urine volume in both groups decreased significantly(all P<0.0S),and the scores and residual urine volume in the observation group were significantly lower than those in the control group(all P<0.05).The inpatient time and catheter indwelling time in the observation group were significantly shorter than those in the control group(both P<0.05).Conclusion:Intradermal needle therapy has an obvious effect in improving symptoms of urinary retention after cervical cancer surgery,and the effect is significantly more persistent than that of simple basic nursing.

Age-specific effectiveness of primary human papillomavirus screening versus cytology in a cervical cancer screening program: a nationwide cross-sectional study

Background:Primary human papillomavirus(HPV)screening is recommended for the detection of cervical intraepithelial neoplasia(CIN)in the general pop-ulation;however,the triage for HPV-positive women remains a challenge.This study aimed to evaluate the age-specific effectiveness of primary HPV screening versus primary cytology screening for identifying optimal strategies for women of different ages.Methods:The dataset of the prevalence round screening was derived from the National Cervical Cancer Screening Program in China.Primary cervical screen-ing protocols included cytology only,HPV testing with cytology triage,and HPV testing with HPV-16/18 genotyping plus cytology triage.The primary outcomes were age-specific detection rate,colposcopy referral rate and positive predictive value(PPV)for CIN2+.Multivariate Poisson regression was used to evaluate the relativeeffectivenessofHPVtestingandcytologyaccordingtoagegroups.TheI 2 statisticwitharandom-effectmodelwasusedtotesttheheterogeneityinrelative effectiveness of HPV testing versus cytology between age groups.Results:This study included 1,160,981 women.HPV testing with HPV-16/18 genotyping plus cytology triage significantly increased the CIN2+detection by 36%(rate ratio[RR]:1.36,95%confidential interval[CI]1.21–1.54)for women aged 35-44 years and by 34%(RR:1.34,95%CI 1.20-1.51)for women aged 45-54 years compared with cytology only.HPV testing with cytology triage had simi-lar CIN2+detection rate compared with cytology only.The PPVs were substan-tially increased for both HPV testing groups.Among women aged 55-64 years old,HPV testing with HPV-16/18 genotyping plus cytology triage increased the colposcopyreferralrateby19%(RR1.19,95%CI1.10-1.29)comparedwithcytology only,butdidnotincreasetheCIN2+detection(1.09,0.91–1.30).Theeffectiveness ofHPVtestingwithcytologytriagedidnotchangeinolderwomen.Thebetween-age-group heterogeneity in the effectiveness was statistically significant for HPV testing with HPV-16/18 genotyping plus cytology triage versus cytology only.Conclusions:Our results suggested that the effectiveness of primary HPV screeningwithdifferenttriagestrategiesdifferedamongagegroups.HPVtesting with HPV-16/18 genotyping plus cytology triage could be used for women aged 35-54 years to detect more lesions,and HPV testing with cytology triage could balance the CIN2+detection and the number of colposcopies for women aged 55-64 years.Longitudinal data including both prevalence and incidence screen-ing rounds are warranted to assess age-specific triage strategies.

Effects of anti-HPV16E6-ribozyme on phenotype and gene expression of a cervical cancer cell line

OBJECTIVE: To investigate the effects of anti-HPV16E6-ribozyme (HRz) on phenotype and gene expression of a cervical cancer cell line. METHODS: HRz was designed by computer programs. HRz's activity was identified by cleavage experiments in vitro. HRz and empty eukaryotic plasmids were transfected into CaSKi cells with lipofectin, then renamed CaSKi-R and CaSKi-P, respectively. The expression of ribozyme in transfected cells was observed by RNA dot blot. The amounts of E6 mRNA in three kinds of cells lines were detected by Northern blot. Cell growth curves and soft agar forming ability were studied. The ability of each cell line to form tumors was assessed in nude mice. Apoptosis rates and expression of c-myc, bcl-2, p53 and Fas were detected by flow cytometry (FCM). Antigens of tumor cells, HLA-1, HLA-2, B7-1 and B7-2 were also detected. NK, LAK, and CD(3)AK cells were induced. Their cytotoxicities were detected in CaSKi-R, CaSKi-P, and CaSKi cells. RESULTS: In vitro cleavage reaction demonstrated that HRz could cleave HPV16E6 mRNA in a site-specific manner. HRz could be expressed stably in transfected CaSKi cells. Northern blot analysis showed that E6 mRNA levels were lower in CaSKi-R than in CaSKi. The growth rate of CaSKi-R was slower than those of CaSKi and CaSKi-P. The soft agar-forming rate of CaSKi-R was lower compared with those of CaSKi and CaSKi-P cells. The ability of CaSKi-R to form tumors in nude mice was also poor. The apoptosis rate of CaSKi-R cells was much higher than those of CaSKi and CaSKi-P. HRz could reduce the expression of E6, c-myc and bcl-2 proteins, and increase the expression of p53 as well. HRz could increase the expression of HLA-2, B7-1 and B7-2 antigens. The cytotoxicity of NK, LAK and CD(3)AK cells was much higher in CaSKi-R than in CaSKi-P and CaSKi cells. CONCLUSION: HRz not only reverses the malignant phenotype of CaSKi cells partially, but also induces apoptosis in the cells, and increases sensitivity of CaSKi cells to immune cells.

Immunomodulatory effects of astragalus polysaccharide on Human peripheral blood mononuclear cells co-cultured with cervical cancer cell lineImmunomodulatory effects of astragalus polysaccharide on Human peripheral blood mononuclear cells co-cultured with cervical cancer cell line

OBJECTIVE:To investigate immunomodulatory effects of Astragalus polysaccharides(APS)on the co-culture of peripheral blood mononuclear cells(PBMCs)with He La cervical cancer cell line.METHODS:To assess the proliferation of PBMCs,carboxyfluorescein succinimidyl ester(CFSE)-labeled PBMCs were co-cultured with He La cells and treated with different concentrations of APS.Supernatants of cell culture were collected for cytokines assay via enzyme-linked immunosorbent assay(ELISA).The impact of APS on the proliferation of PBMCs,induction of regulatory T cells(Tregs),and myeloid-derived suppressor cells(MDSCs)was carried out by flow cytometry.RESULTS:It was observed that APS could increase the proliferation of PBMCs co-cultured with He La cells(P<0.05).However,APS had no significant effects on the induction of Tregs and MDSCs in the co-culture assay(P>0.05).Furthermore,ELISA results demonstrated that APS could decrease IL-10 and TGF-βconcentration(P<0.05).CONCLUSION:The above-mentioned characteristics showed that APS might be able to modulate immune responses and improve anti-tumor effects through increasing the proliferation of PBMCs and decreasing inhibitory cytokines secretion as critical mediators of immune suppression in the tumor microenvironment.