1.Evaluation of the application of folate receptor mediated (FRD) multifunctional acetic acid white solution inspection with liquid based cytology screening for cervical cancer
Cui DENG ; Jiali KANG ; Wenyan JIANG ; Miaoling NIE ; Xiaohui HUANG
Journal of Chinese Physician 2014;16(8):1064-1067
Objective To explore the folate receptor mediated (FRD) check and multi-function acetic acid white solution liquid based cervical cytology (TCT) application value in cervical cancer screening.Methods A total of 602 cases of patients was tested with FRD multi-function acetic acid white solution check,and TCT and cervical biopsy pathology examination.With the used of histopathological results as the gold standard,FRD multi-function acetic acid white analysis was compared with the TCT screening inspection results.Results For a total 602 patients with TCT screening,the positive rate was 21.8% (131/602),including 36 cases of CIN Ⅰ level,41 cases of CIN Ⅱ level,24 cases of CIN Ⅲ level,and 30 cases of cervical invasive carcinoma.For the FRD multifunction white acetate solution screening,its positive rate was 23.8% (143/602).No statistically significant difference was found between TCT and FRD screening (P > 0.05).The missed diagnosis rate of FRD multi-function white acetate solution screening was 2.6% in inflammation,and 21.1% in cervical invasive cancer,and 3.8 % in CIN.The missed diagnosis rate of TCT screening was 7.2% in inflammation,5.3% in CIN Ⅰ,4.9% in CIN Ⅱ,and 58.6% in CIN Ⅲ]; whereas,its detection coincidence rate was 100% in squamous cells carcinoma (SCC) and adenocarcinoma (AC).FRD multi-function acetic acid white solution screening had a sensitivity 80.92%,specificity 92.14%,positive predictive value 74.13%,and negative predictive value 95.59%.TCT examination had a sensitivity 90.84%,specificity 90.23%,positive predictive value 72.12%,and negative predictive value 97.25%.No significant difference was found between FRD and TCT methods (P > 0.05).Conclusions FRD and TCT methods were both efficient in screening and evaluation for cervical lesions and cervical cancer.Because FRD method is limited in the deep tube for examination of cervical lesions; it cannot completely replace the TCT examination.However,FRD method is reliable,economic,and simple operation; it is suitable for primary hospitals census of cervical cancer
2.Expression of epidermal growth factor receptor of cervical carcinoma and its correlation with prognosis
Linghong DENG ; Shuyu FENG ; Jiali KANG ; Yanna ZHANG ; Qiaoer LIN ; Miaoling NIE ; Haiyan CHENG ; Jinrui SUN
Clinical Medicine of China 2009;25(6):590-592
Objective To study the expression of epidermal growth factor receptor (EGFR) in cervical car-cinoma and cervical intraepithelial neoplasia(CIN),and to elucidate its relation with the genesis, infiltration, metas-tasis and prognosis of cervical carcinoma. Methods EGFR was determined by means of S-P immunohistochemistry in tissue of 100 cases of cervical carcinoma,60 cases of CIN and 40 cases of controls. Results The overexpression rates of EGFR were 0% (0/40), 51.67% (31/60),78.00% (78/100), respectively in normal cervical epithelium, CIN and cervical tumor tissues. The overexpression rate of EGFR was significantly higher in cervical tumor tissue than in control group(P<0.01). The overexpression of EGFR didn't demonstrate significant association with clinical staging, tumor size, pathological type, differentiation, cervical invasion depth, cervical canal invasion, lymphnode me-tastasis or the prognosis of cervical neoplasm (P>0.05). Conclusion Overexpression of EGFR is worsened with the severity of cervical lesion, suggesting that overexpression of EGFR is correlated with the genesis of cervical neo-plasms,which may be a valuable biological indicator of cervical carcinoma,but is not correlated with clinical patho-logical features and prognosis of cervical carcinoma.
3.Effect and mechanism of gene therapy of lentivirus mediated RhoA shRNA on ovarian cancer xenograft in vivo
Wenyan JIANG ; Jiali KANG ; Xiaoxia WANG ; Wenjuan YANG ; Miaoling NIE ; Cong ZHOU
Chinese Journal of Obstetrics and Gynecology 2013;48(10):778-783
Objective To investigate treatment effects of lentivirus mediated RhoA short hairpin RNA(shRNA) on xenograft tumor of ovarian cancer in nude mice in vivo and the underlying mechanism.Methods Human ovarian cancer cell line HO8910 were inoculated to establish subcutaneous xenograft model of human ovarian cancer.Tumor-bearing nude mice were assigned randomizely to three groups:LentiRhoA-sh group,Lenti-negative control (NC) group and phosphate buffered saline (PBS) group.lentivirus mediated RhoA shRNA,negative control lentivirus and PBS were respectively injected in the three groups.Effects of treatment were observed by tumor growth curve,tumor volume,tumor weight,and tumor inhibition rate.Xenograft tissues and liver,spleen,lung,and renal tissues were examined by hematoxylin and eosin (HE) staining or were detected by streptavidin-perosidase (SP)immunochemical method.The changes of RhoA gene expression in xenograft tissues after lentivirus mediated RhoA shRNA treated were also detected by real-time qPCR,immunochemistry and Western blot assay.Cell apoptosis in xenograft tissues were examined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL) method and apoptotic index (AI) were counted.Results Compared with Lenti-NC group and PBS group,the growth speed of xenograft in Lenti-RhoA-sh group delayed significantly after injection 9 days(P < 0.01).Tumor volume (338 ± 114) mm3 decreased significantly in the Lenti-RhoA-sh group when compared with those in Lenti-NC group(1190 ± 332)mm3 and PBS group (1101 ± 396) mm3 (P < 0.01).Tumor weight (0.23±0.11)g decreased significantly in the Lenti-RhoA-sh group when compared with Lenti-NC group (0.79 ± 0.19)g and PBS group (0.74 ± 0.17)g (P < 0.01).Real-time qPCR result shown that the expression of RhoA mRNA (0.30 ± 0.05) decreased significantly in the Lenti-RhoA-sh group compared with Lenti-NC group (0.95 ±0.06) and PBS group(1.00 ±0.11 ; P <0.01).Western blot result showed that the expression level of RhoA protein decreased significantly in the Lenti-RhoA-sh group (0.14 ± 0.06) compared with those in Lenti-NC group(0.78 ± 0.14) and PBS group (0.75 ± 0.13;P < 0.01).TUNEL staining displayed that AI significantly increased in the Lenti-RhoA-sh group (20.9 ± 3.4) % compared with those in Lenti-NC group(5.2 ±±2.0)% and PBS group(6.0 ±2.1)% (P <0.01).Conclusion Lentivirus mediated RhoA shRNA may be effectively down-regulate of the expression of RhoA,inhibit the growth of subcutaneous xenograft tumor of ovarian cancer in nude mice by increasing the cell apoptosis.
4.Screening of targeting peptides for highly metastatic human ovarian cancer cells and their effect on the biological behavior of ovarian cancer cells.
Cong ZHOU ; Jiali KANG ; Xiaoxia WANG ; Miaoling NIE ; Wenyan JIANG
Chinese Journal of Oncology 2014;36(8):565-570
OBJECTIVETo explore the effect of short peptides specifically binding to highly metastatic human ovarian cancer HO8910PM cells and their effect on the biological behavior of ovarian cancer cells.
METHODSThe phage-displayed peptide library was used to isolate the peptides binding and internalizing into the HO8910PM cells. Positive phage clones were characterized with DNA sequencing and bioinformatics analysis. The positive phage clones specifically bound to HO8910 cells were validated with immunofluorescence detection and enzyme-linked immunosorbent assay (ELISA). Furthermore, selected peptides were investigated for their cancer-related functions, including cell adhesion, spreading, motility, and invasion in vitro and in nude mice in vivo. The apoptotic index was detected by TUNEL assay, and VEGF expression by immunohistochemistry.
RESULTSAfter 4 rounds of screening, apparent enrichment of phages was observed on the HO8910PM cells. ELISA assay showed that among the randomly selected 20 phage clones, 12 can specifically bind to HO8910PM cells. Immunofluorescence assay also showed that the selected positive phage clones can specifically bind to HO8910PM cells. The adherence test showed that the adherence rates of HO8910PM-peptide20, HO8910PM-peptide16 and HO8910PM cells were 49.0%, 96.8% and 100.0%, respectively. There was a significant difference between the cell adherence rates of HO8910PM-peptide20 and HO8910PM cells (P < 0.05). The peptide20 read as "THRVHLH" was a positive peptide and showed preferential binding to targeted cells. The peptide20 effectively inhibited tumor growth and metastasis in the nude mice, and the positive rates of VEGF protein in the tumor tissue of experimental, negative control and blank mice were 21.2%, 81.4% and 85.7%, respectively, showing that the positive rate of VEGF protein in the experimental group was significantly lower than that in the negative control and blank groups (P < 0.01), and the apoptotic index (AI) of the experimental group was (18.21 ± 2.49)%, significantly higher than the (3.76 ± 1.77)% in the negative control group and the (4.78 ± 1.57)% in the blank group (P < 0.01).
CONCLUSIONSA novel short peptide able to specifically bind to highly metastatic human ovarian cancer cells is successfully screened. It can effectively inhibit the growth, invasion and metastasis of ovarian cancer cells, and provides an ideal vector in targeted drug therapy for ovarian cancer.
Animals ; Base Sequence ; Cell Adhesion ; Cell Line, Tumor ; Cell Movement ; Enzyme-Linked Immunosorbent Assay ; Female ; Genetic Vectors ; Humans ; Immunohistochemistry ; Mice ; Mice, Nude ; Molecular Targeted Therapy ; methods ; Neoplasms, Glandular and Epithelial ; metabolism ; Ovarian Neoplasms ; metabolism ; Peptide Library ; Peptides ; metabolism