1.Inhibition on the proliferation of human endometrial cancer cells by RNAi inhibiting HMGB1 gene expression and its possible molecular mechanism
Journal of Central South University(Medical Sciences) 2014;(7):673-680
Objective: To investigate the effect of HMGB1 small interfering RNA (siRNA) on the proliferation, cell cycle and apoptosis of human endometrial cancer cell line HEC-1A, and itspossible molecular mechanism. Methods: Lentivirus vector with HMGB1 shRNA was constructed and infected the endometrial cancer cell line HEC-1A. After viral infection for 72 h, real time PCR and Western blot were performed to investigate HMGB1 mRNA and protein expression. The cell proliferation was determined with methyl thiazolyl tetrazolium (MTT) method. Flow cytometry was performed to analyze the cell cycle progression of propidium iodide (PI)-stained HEC-1A cells and the apoptotic rate of annexinV/PI-stained cells. Western blot was used to detect the protein expression of AKT, pAKT and CyclinD1. Results: Lentivirus vector with HMGB1 shRNA inhibited the mRNA (P<0.05) and protein (P<0.01) expression of HMGB1 in the cell line HEC-1A. The MTT assay demonstrated that HMGB1 knockdown signiifcantly reduced the cell proliferation. FCM results showed that HMGB1 knockdown significantly resulted in the disruption of the cell cycle at G0/G1 phase and the induction of apoptosis. hTe apoptotic rate was (17.89±0.23)%, (4.69±0.20)% and (4.62±0.17)% in the HMGB1 knockdown group, the blank group and the negative group respectively. There was signiifcance difference between the 3 groups (P<0.01). hTe protein expressions of pAKT and cyclinD1 were down-regulated atfer the HMGB1 knockdown for 72 h. Conclusion: Knockdown of HMGB1 expression can significantly inhibit the proliferation and induce the cell cycle arrest and apoptosis in the endometrial cancer cell line HEC-1A. PI3K/AKT pathway and down-regulation of the protein expression of cyclinD1 may be involved in its therapeutic mechanism.
2.Influence of CD146-siRNA on epithelial-mesenchymal transition in human ovarian cancer cell line A2780
Journal of Chinese Physician 2014;16(10):1374-1376
Objective To investigate the influence of CD146-siRNA on epithelial-mesenchymal transition in human ovarian cancer cell line A2780.Methods The influence of loss of CD146 on the mRNA and protein expressions of E-cadherin,vimentin,and fibronectin was evaluated by real-time polymerase chain reaction (RT-PCR) and Western blotting.Results Compared to non-transfected (control group) and those transfected with RNAi-NC (RNAi negative control group),the mRNA and protein expressions of Ecadherin were significantly increased in A2780 cell lines transfected with RNAi-CD146.The loss of CD146 expression down-regulated the mRNA and protein expression of vimentin and fibronectin(P < 0.05).Conclusions CD146-siRNA can suppress epithelial-mesenchymal transition in human ovarian cancer cell line A2780.
3.The expression and significance of high mobility group protein box1 in ovarian endometriosis tissue
Journal of Chinese Physician 2013;15(10):1349-1352
Objective To investigate the expression of high mobility group protein box1 (HMGB1)in ovarian endometriosis tissue and to provide new idea for early diagnosis,treatment and prognosis of endometriosis.Methods The levels of HMGB1 in ectopic endometrium and eutopic endometrium of 69 patients with ovarian endometriosis and endometrium of 69 cases with non-endometrioses were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western-blotting.Results HMGB1 was expressed in all the three groups (ectopic endometrium eutopic endometrium endometrium with non-endometrioses).The relative expression level of HMGB1 mRNA was 0.54383 ± 0.0565,0.3016 ± 0.0614,and 0.1536 ±0.0444,respectively (F =377.923,P < 0.0001).The relative of HMGB1 protein was 0.83990 ±0.12869,0.40100 ±0.16425,and 0.19986 ± 0.08162,respectively (F =185.625,P =0.0000).The expression level of HMGB1 was related to clinical stages (t =10.28,P <0.01) ; however,it was not obviously related to age,the diameter of lump,and course of disease (P > 0.05).Conclusions Over expression of HMGB1 in ovary endometriosis indicates that HMGB1 plays a significant role in occurrence and development of ovary endometriosis.The expression of HMGB1 in endometriosis eutopic endometrium was much higher than that of non-endometrioses endometrium,which supports the eutopic endometrium determinism.The overexpression of HMGB1 was consistent with clinical stage,which proves that HMGB1 maybe be tightly associated with the severity of the disease and to likely become biomarker.
4.Causes and Prevention of the Uterus Repture in an Outlying District of Hunan Province
Weishe ZHANG ; Zhaodi WU ; Jiajie WU
Journal of Chinese Physician 2001;0(08):-
Objective To study the causes of the uterus rupture in an outlying district and prevent measures.Methods 17 cases of the uterus rupture in Yongshun county hospital were retrospectively analysized.Results Among 17 cases of uterus rupture,the average antenatal care was 0 4 times every pregnant women,bleeding volume were 1100 ml,12 cases of shock(70 6%) ,5 cases forced uterectomy(29 4%), 16 cases of perinatal death(94%). According to stages of taking place : ⑴15 cases had taken place during laboring without appointing inpatient ,13 patients giving birth at home while 2 patients in hospital , average total stages of labor 29.4h, abnormal fetal positions 14 cases, including 3 cases of primipara and 12 cases of multipara. ⑵ 2 cases of spontaneous uterus rupture occurred during pregnancy who have with the cesarean section history performed in small clinic in the country side.Conclusions In an outlying district, main causes of uterus rupture:⑴ Ignored antenatal care. ⑵ Producing at home so that dystocia were not dealt with properly during stage of labor, another was women with prolong labor treated incorrectly . ⑶ The Spontaneous rupture during pregnancy were close related to the history of the classical cesarean section .To prevent the uterus rupture should be enhanced prenatal care and examination forced pregnant women delivery in hospital, standarding medical practice of performing operation, so doing could sharply decrease the pregnant uterus rupture in an outlying district.
5.Interactions of anticancer drugs and miRNA targets
Jiajie ZHANG ; Guangfa WANG ; Shuguang WU
Chinese Pharmacological Bulletin 2003;0(11):-
miRNA are large class of small noncoding RNAs that play important roles in the development of various diseases such as cancers.This review summarizes the miRNA biogenesis and related enzymes or cofactors,the alteration of miRNA expression induced by antitumor drugs as well as the interaction of these drugs and miRNA.The paper also predicts the future research aspects for miRNA,including the functional studies of miRNAs,the drug screening based on miRNA and the regulatory mechanisms of miRNA expression by drugs.
6.Study of Systemic Lupus Erythematosus (SLE) Treatment and Disease Activity
Hongfu XIE ; Jie LI ; Jiajie WU
Journal of Chinese Physician 2000;0(11):-
Objective To investigate the changes of disease activity and clinical significance in the course of treatment in patients with SLE.Method 286 cases of SLE were reviewed and compared the changes of SLEDAI scores in different disease duration.Results The SLEDAI scores of patients whose first treatment courses less than 1 month and 1 to 3 months were significantly lower than those patients whose were 4 to 6 months and more than 6 months. After treatment for 2 months to 3 years, the SLEDAI scores were not correlated with cumulated dosage of corticosteroids.Conclusions For the patients of short first treatment course, the treatment could relieve SLE disease activity rapidly and effectively to some extent; while for the patients whose first treatment courses were relatively long ,the relif of disease activity was relatively slow. After treatment for 2 to 3 months, the disease of SLE patients was more active than other periods, and it was inclined to produce visceral damage. As mentioned above ,we should pay attention to this phenomenon.
7.Study of Cumulative Organ Damage of Systemic Lupus Erythematosus (SLE) and It's Affecting Factors
Hongfu XIE ; Jie LEE ; Jiajie WU
Journal of Chinese Physician 2001;0(01):-
Objective To investigate the cumulative organ damage of systemic lupus erythematosus (SLE) and It's affecting factors.Methods 162 cases of SLE patients were reviewed and evaluated in the cumulative organ damage of them . At the same time, It's affecting factors were analysed by multifactorial analysis.Results The cumulative organ damage of SLE was obviously correlated with the first time of treatment, treatment with CTX ,the numbers of recurrence rate and level of complements;but were not correlated with disease duration and cumulative dosage of corticosteroid.Conclusions The cumulative organ damage was one of the evaluating factors of SLE disease, and it's occurence and development were affected by multiple factors .So, the patients should be treated with hormone and control the beneficial factors to protect organs,such as,observation on complemets level,high dosage cyclophosphamide pulse treatment etc.
8.Correlation with expression of HMGB1 mRNA and clinicopathological features of endometrial carcinoma
Rixue XIE ; Jiajie WU ; Ying XIE ; Meifang XIAO ; Cuixiang ZHOU
Journal of Chinese Physician 2011;13(1):41-43,48
Objective To investigate the expression of HMGB1 mRNA in endometrial carcinoma and its correlation with clinicopathological features of endometrial carcinoma. Methods Semi-quantitative RT-PCR method was used to detect the expression level of HMGB1 gene in 56 cases of endometrial carcinoma and 20 cases of normal endometrium. Results The expression of HMGB1 mRNA in endometrial carcinoma (0. 3512 ± 0. 0985 ) was significantly higher than that in normal endometrium (0. 2208 ± 0. 0170 ).There was a significant difference in the two groups( P <0. 05 ). Grouping by clinicopathological features,the expression of HMGB 1 mRNA in endometrial carcinoma had a correlation with clinical-surgical stage ( P<0. 05), metastasis of lymph nodes ( P < 0. 05), and depth of myometrial invasion ( P < 0. 05 ). Conclusion The high level of HMGB1 mRNA indicated that HMGB1 might play an essential role in the genesis, growth, invasion and metastasis of endometrial carcinoma.
9.Significance of expression of HMGB1 and Smac/DIABLO protein in epithelial ovarian cancer
Cuixiang ZHOU ; Jiajie WU ; Qiao YU ; Rixue XIE
Journal of Chinese Physician 2011;13(11):1455-1458,1463
ObjectiveThe purpose of this study was to investigate the clinical significance of HMGB1 and Smac/DIABLO protein expression in epithelial ovarian cancer.MethodsImmunohistochemistry (IHC) and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining were used to detect the protein expression of HMGB1,Smac/DIABLO and cell apoptosis in 45 epithelial ovarian cancers by tissue microarray.ResultsThe protein expression of HMGB1 in epithelial ovarian cancers was significant higher than benign cancers and normal ovarian tissues( P <0.05).The protein expression of HMGB1 was related to the lymph node metastasis,which had a negative correlation with the cell apoptosis index ( rs =-0.583,P =0.000).The protein expression of Smac/DIABLO in epithelial ovarian cancers was significantly lower than benign cancers and normal ovarian tissues( P <0.05) ; the protein expression of HMGB1 in epithelial ovarian cancers had a negative relationship with protein expression of Smac/DIABLO( rs =-0.40,P =0.006).ConclusionsThe over-expression of HMGB1 in epithelial ovarian cancers may play an important part in the metastasis of the epithelial ovarian cancers.The over-expression of HMGB1 and the low-expression of Smac/DIABLO may take part in the occurrence and development of the epithelial ovarian cancers through abnormal cell apoptosis.
10.Over-expression and suppression of HMGBI affect proliferation and invasion of endometrial carcinoma and underlying mechanisms
Jing WANG ; Yuqi GAO ; Xinyi LIU ; Jiajie WU
Journal of Central South University(Medical Sciences) 2017;42(7):769-775
Objective:To investigate effects of over-expression and suppression of HMGB1 on proliferation and invasion of endometrial carcinoma HEC-1A cell and underlying mechanisms.Methods:Over-expression or silence of HMGB 1 in HEC-1A cell lines were established by lentiviral vector containing HMGB 1 recombinant plasmid or by HMGB 1 shRNA,respectively.Cell counting kit-8,Transwell,and wound healing assay were used to analyze proliferation,invasion,and migration of HEC-1A cells,respectively.Western blot and reverse transcription-PCR (RT-PCR) were used to detect the expression of NF-κB,VEGF,and matrix metalloproteinase 2 (MMP2) in the cells.Results:Over-expression of HMGB1 promoted the proliferation,invasion,and migration of HEC1A cell,and up-regulated NF-κB,VEGF,and MMP2 expressions,while suppression of HMGB1 inhibited the proliferation,invasion,and migration of HEC-1A cells and down-regulated NF-κB,VEGF,and MMP2 expressions.Conclusion:HMGB1 plays an important role in proliferation,invasion,and migration of HEC1A cell via NF-κB/VEGF/MMP2 pathway.HMGB 1 might be a potential target for endometrial carcinoma therapy.