Expressions and Significance of Livin, Survivin, Pak-1 and Caspase-7 in Different Gastric Mucosal Lesions / 胃肠病学

Background: Livin, survivin and Pak-1 are all related to the occurrence and development of gastric cancer. Livin and survivin play their roles by inhibiting the activity of caspase-7. Aims: To investigate the expressions and significance of livin, survivin, Pak-1 and caspase-7 in different gastric mucosal lesions. Methods: A total of 45 cases of gastric cancer and paracancerous tissue, 45 chronic atrophic gastritis with intestinal metaplasia, 45 chronic non-atrophic gastritis from Jan. 2015 to Dec. 2019 at the Second Affiliated Hospital of Baotou Medical College were collected. Immunohistochemistry was used to detect the expressions of livin, survivin, Pak-1 and caspase-7, and their correlations with clinicopathological features of gastric cancer patients were analyzed. Results: Compared with chronic non-atrophic gastritis group and paracancerous group, the positivity expression rates of livin, survivin and Pak-1 in intestinal metaplasia group were significantly increased (P<0.05), while caspase-7 was significantly decreased (P<0.05). Compared with intestinal metaplasia group, the positivity expression rates of livin, survivin and Pak-1 in gastric cancer group were significantly increased (P<0.05), while caspase-7 was significantly decreased (P<0.05). Expressions of livin, survivin, Pak-1 and caspase-7 were correlated with the differentiation degree, TNM stage, depth of infiltration and lymph node metastasis in patients with gastric cancer (P<0.05). Conclusions: Livin, survivin, Pak-1 and caspase-7 play important roles in the occurrence and development of gastric cancer.

Suppressive effect of Tet-on mediated livin RNA interference on growth of lung carcinoma xenegraft in a rat model / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate he effect of tetracycline- (Tet-on) mediated livin RNA interference on growth of lung carcinoma xenegrafts, and find a better regulatory way to interfere the development on lung cancer. Methods: livin shRNA lentiviral vectors were constructed; and the lung cancerA549 cells were subcutaneously injected into right upper back of nude mice to establish xenegraft model. The livin shRNAlentiviral vectors were injected into xenografts to interfere the expression of livin, then tetracycline was injected intraperitoneally for the induction. The suppressive effect of Tet-on mediated livin RNA interference efficiency was investigated and lung cancer xenograft development was observed. Results: After the induction with Tet-on, livin gene expression was significantly inhibited by livin shRNAcompared with the control group and Tet-on-NC group; the xenograft volume in Tet-on- livin shRNAgroup was significantly smaller than that in control group and Tet-on-NC group ([5.31±0.86]g vs [8.22±0.63]g and [7.17±0.54] g, P<0.05). Moreover, little body toxicity was observed and no nude mice died in this study. Conclusion: The Tet-on mediated livin shRNA could suppress the growth of lung cancer development with good targeting and controllable characteristics, which might provide a potent tool for treating lung cancer with livin protein as target.

Expression of PDCD4 and apoptosis inhibitor Livin in triple negative breast cancer tissues and its relationship with prognosis / 重庆医学

Objective To observe and analyze the expression of programmed cell death 4 (PDCD4) gene and apoptosis inhibitor Livin in triple negative breast cancer (TNBC) tissues and its relationship with prognosis.Methods One hundred cases of TNBC tumor tissue,50 cases of adjacent carcinoma tissue,50 cases of normal breast tissue were selected as the research data.The immunohistochemical technique was applied to detect and compare the expression positive rates of PDCD4 and Livin protein in three kinds of tissues.The patients were followed up.The overall survival (OS) and the progression free survival (PFS) were observed and compared.Results The expression positive rate of PDCD4 in TNBC tissue was significantly lower than that in adjacent carcinoma tissue or normal breast tissue,the differences were statistically significant (x2=26.613,32.000,P＜0.05).The expression was correlated with the clinical pathological features of tumor size,lymph node metastasis,clinical stage,axillary lymph node metastasis and cancer embolus (x2=26.936,13.210,22.774,27.463,5.803,P＜0.05);the expression positive rate of Livin protein in TNBC tissue was significantly higher than that in adjacent carcinoma tissue or normal breast tissue and the expression positive rate of Livin protein in adjacent carcinoma tissue was significantly higher than that in normal breast tissue,the differences were statistically significant (x2 =14.614,57.353,19.048,P＜0.05).The expression was correlated with the clinical pathological features of lymph node metastasis,clinical stage,axillary lymph node metastasis and cancer embolus (x2 =10.788,6.160,27.350,8.914,P＜0.05);OS,PFS in the patients with PDCD4 negative expression were significantly lower than those in the patients with PDCD4positive expression.OS,PFS in the patients with Livin positive expression were significantly lower than those in the patients with Livin negative expression,the above differences were statistically significant (x2 =23.931,19.163,22.649,17.213,P＜0.05).OS in the TNBC patients was correlated with age (RR=1.405),clinical stage (RR =2.897),tumor diameter (RR=2.722),axillary lymph node metastasis (RR=2.516),vascular invasion (RR=3.020),PDCD4 Expression (RR=1.752) and Livin expression (RR=2.051) (P＜0.05).PFS in the patients was correlated with clinical stage (RR =2.756),axillary lymph node metastasis (RR =2.437),PDCD4 expression (RR =1.649) and Livin expression (RR=1.804) (P＜0.05).Conclusion The PDCD4 low expression and Livin protein over-expression exist in TNBC tissues.Their abnormal expressions are correlated with the clinicopathological features of tumor and the prognosis of patient,and could be used as the auxiliary indexes in evaluation of progression and prognosis of TNBC.

Clinical expression of P53,Livin and PARP in the epithelial ovarian cancer and its impact in the chemotherapy resistance and clinical prognosis / 中国生化药物杂志

Objective To explore the clinical expression of P53, Livin and PARP in the epithelial ovarian cancer and its correlation with the chemotherapy resistance and clinical prognosis.Methods 74 specimen of epithelial ovarian cancer confirmed from January 2009 to June 2011 in our gynecology department were selected.During the follow-up visit, the subjects were divided into chemotherapy sensitivity group and chemotherapy resistance group according to the recurrence cases, the clinical expression and survival rate for two groups were compared, the influence factors of survival time were analyzed.Results The positive rate of P53, Livin and PARP for chemotherapy sensitivity group was 47.1%, 56.9%and 52.9%;the positive rate for chemotherapy resistance group was 73.9%, 95.7% and 95.7%,the diyforences were significant(P<0.05).After 1, 3 and 5 years of treatment, the survival rate for chemotherapy sensitivity group was 100.0%, 82.4% and 66.7%,The survival rate for chemotherapy resistance group was 87.0%, 26.1% and 8.7%,the diyforences were significant(P<0.05).Based on the Cox regression model, the influence factors of the patient's age, pathological differentiation degree, clinical staging and chemotherapy sensitivity were introduced.It was known that the patient's survival time was greatly influenced by clinical staging and chemotherapy sensitivity (P<0.05).Conclusion For patients with epithelial ovarian cancer, the expression of P53, Livin and PARP is correlated with chemotherapy resistance.Therefore, the clinical effect is predictable, for patients with higher expression, the personalized therapy can improve the patient's prognosis.

Silencing the livin gene enhances the cytotoxic effects of anticancer drugs on colon cancer cells

PURPOSE: Livin is associated with drug response in several cancers. The aim of this study was to investigate the effect of silencing the livin gene expression on anticancer drug response in colorectal cancer. METHODS: siRNA was transfected at different concentrations (0, 10, and 30nM) into HCT116 cells, then cells were treated with either 5-fluorouracil (FU)/leucovorin (LV) or oxaliplatin (L-OHP)/5-FU/LV. Cellular viability and apoptosis were evaluated following silencing of livin gene expression combined with treatment with anticancer drugs. RESULTS: Livin gene expression was effectively suppressed by 30nM siRNA compared with control and 10nM siRNA. The 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay showed that proliferation was effectively inhibited in cells treated with a combination of both siRNA and an anticancer drug, compared to cells treated with siRNA-Livin or anticancer drug alone. In particular, the combination of 30nM siRNA and L-OHP/5-FU/LV resulted in a 93.8% and 91.4% decrease, compared to untreated control or L-OHP/5-FU/LV alone, respectively. Cellular proliferation was most effectively suppressed by a combination of 30nM of siRNA and L-OHP/5-FU/LV compared to other combinations. CONCLUSION: siRNA-mediated down-regulation of livin gene expression could significantly suppress colon cancer growth and enhance the cytotoxic effects of anticancer drugs such as 5-FU and L-OHP. The results of this study suggest that silencing livin gene expression in combination with treatment with anticancer drugs might be a novel cancer therapy for colorectal cancer.

Effect of livin-modified BM-MSCs transplantation on cardiac function fol-lowing acute myocardial infarction in a rat model / 中国病理生理杂志

[ ABSTRACT] AIM:To study the effect of livin gene-modified bone marrow mesenchymal stem cells ( BM-MSCs) transplantation on the cardiac function following acute myocardial infarction in a rat model and the expression of livin , caspase-3, caspase-7 and caspase-9 in the livin gene-modified BM-MSCs.METHODS: The MSCs were obtained by the whole bone marrow culture method , and the apoptosis of the MSCs after infection with adenovirus vector carrying enhanced green fluorescent protein ( EGFP) gene and livin recombinant vector ( rAd-livin) were detected by flow cytometry .The ex-pression of livin, caspase-3, caspase-7 and caspase-9 was detected by Western blot .After permanent left anterior descend-ing artery occlusion , the rats were randomized to receive intramyocardial injection of DMEM without cells ( vehicle group ) , or containing MSCs ( MSCs group ) , MSCs ( EGFP ) ( rAd-control/MSCs group ) or MSCs ( livin ) ( rAd-livin/MSCs group).Left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), the maximum in-creased rate of left ventricular pressure ( -dp/dtmax ) and the maximum decline rate of left ventricular pressure ( +dp/dtmax ) were recorded for evaluating the cardiac functions .RESULTS: The apoptosis of rAd-livin/MSCs was significantly decreased as compared with MSCs and rAd-control/MSCs (P<0.05).Meanwhile, the expression of caspase-3, caspase-7 and caspase-9 was significantly downregulated as compared with the other 2 groups ( P<0.05 ) .The cardiac function in rAd-livin/MSCs group was significantly improved as compared with DMEM group , and those in the other 2 groups got the similar results, but the function in rAd-livin/MSCs group was better improved .Meanwhile, the number of surviving cells in rAd-livin/MSCs group was significantly improved as compared with the other 2 groups .CONCLUSION:The apoptosis of MSCs is decreased after rAd-livin transfection, and the expression of caspase-3, caspase-7 and caspase-9 is also significant-ly downregulated while the expression of livin is significantly upregulated .Transplantation of livin-modified BM-MSCs by lentiviral vector results in better prognosis for treating myocardial infarction by enhancing cell survival .

Effect of Livin on Proliferation and Multi-drug Resistance of Lung Cancer Cells A549 / 中国医科大学学报

Objective To study the effect of the inhibitor of apoptosis protein,Livin on proliferation and multi?drug resistance of lung adenocarcino?ma cells A549. Methods A549 cells were transfected with the eukaryotic expression vector pcDNA3.1?Livin. A549 cell clone with stable expres?sion of Livin was obtained through G418 screening. Expressions of Livin mRNA and protein in the transfected cells were respectively measured by re?verse transcription polymerase chain reaction(RT?PCR)and Western blot. The distribution of cell cycle phase was determined using flow cytometry. The level of P?gp mRNA and protein in A549 cells transfected with pcDNA3.1?Livin was detected by RT?PCR and Western blot. The analysis of multi?drug resistance of A549 treated with different chemotherapeutics was performed by MTT. Results The mRNA and protein expressions of Liv?in were both significantly increased in the transfected A549 cells. The flow cytometry analysis showed there was higher percentage of S phase and low?er percentage of G0/G1 phase in A549 cells transfected with pcDNA3.1?Livin. Compared with control groups,the expression of P?gp mRNA and pro?tein was increased in A549 cells transfected with pcDNA3.1?Livin,which showed a higher drug resistance and lower sensitivity to chemotherapic drugs such as ADM,MTX,CTX,and DDP(P<0.05). Conclusion Overexpression of Livin could enhance the proliferation of A549 cells,and high expression of P?gp caused by Livin could serve as one of the causes for multi?drug resistance in lung adenocarcinoma against chemotherapies.

Livin and Urological Malignancy, related research and progress / 医学研究生学报

Oncogenesis is a sophisticated process which is polygenic , multi-stage,and multi-step.Livin as an apoptosis inhibi-ting factor , is one of the members of apoptosis inhibiting factor family , it expresses highly in many tumor tissues , and is closely related to the occurrence and development of urological tumor .Tumor growth can be inhibited by interfering the expression of Livin in tumor tis-sue,which will reduce the resistance to apoptosis and increase the rate of cell apoptosis .In recent years , more and more attention was paid to the studies of Livin on gene targeting therapy .Further research of Livin might provide new ideas for the early diagnosis , clinical treatment and evaluation of prognosis of tumor .

Effects of siRNA targeting livin gene on proliferation and apoptosis of nasopharyngeal carcinoma cell line CNE-2Z / 中国免疫学杂志

To observe the effect of livin gene-specific siRNA interference on proliferation and apoptosis of nasopharyngeal carcinoma (NPC) cell line CNE-2Z.Methods: siRNA expression vectors pGPU6/GFP/Neo-livin were transfected into NPC cell line CNE-2Z by using Lipofectamine 2000.The expressions of livin mRNA and protein were detected by semi-quantitative reverse transcriptase-polymerase chain reaction ( RT-PCR) and Western blot.The changes of caspase-3 activity were assessed by kinase semi-quantitative activity test.The proliferation activity and apoptosis of CNE-2Z cells were examined by MTT and flow cytometry respectively.Results:The expression levels of livin mRNA and protein in pGPU 6/GPF/Neo-livin transfected CNE-2Z cells were significantly lower than those in untreated and pGPU 6/GFP/Neo-shNC ( control non-target siRNA ) transfected cells with the expression inhibitory rate of 64.38% and 61.43% respectively.The caspase-3 activity and the apoptotic rate of experimental group cells were increased obviously.The growth of CNE-2Z cells was inhibited by siRNA recombinant expression vector transfection.Conclusion:siRNA targeting livin gene inhibits proliferation and induce apoptosis of CNE-2Z cells.

Effects of different types of music on neuronal apoptosis of hippocampus of brain tissue of aged rats / 中国免疫学杂志

Objective:To explore the effects of different types of music on Caspase-3 and Livin expression levels of the hippocampus of brain tissue of aged rats.Methods: Healthly elderly male Wistar rats 40 were randomly divided into :control group , Mozart music group ,Liangzhu music group and rock music group ,n=10;water maze training every morning while music intervention 2 h,afternoon pure music intervention 2 h,continuous 7 d,the eighth days ,the rats were sacrificed and taking blood and brain tissue ,by ELISA in serum and brain tissue Caspase-3 and Livin protein contents change;using immunohistochemistry assay in rat brain tissue Caspase-3 and Livin protein levels;real-time PCR detection of rat brain tissue changes in gene level of Caspase-3 and Livin.Results:Compared with the control group ,Mozart music group ,Liangzhu music rats the protein content ,protein level and gene level of Caspase-3 of serum and brain tissue were lower statistically significant ( P0.05 );while rock music group the protein content,protein level and gene level of Caspase-3 and Livin of brain tissue were not significantly difference (P>0.05).Conclusion:Mozart music group and Liangzhu music group by reducing serum and brain tissue Caspase -3 and increased Livin , to inhibit the apoptosis of the body′s nerve cells,which play a neuroprotective effect;the rock music group in the present study ineffective.

Expression of livin and beclin1 in epithelial ovarian cancer and its rela-tions with tumor invasion and metastasis / 中国现代医生

Objective To explore the expression of Livin and Beclin1 in epithelial ovarian cancer and their relations with tumor invasion and metastasis. Methods A total of 65 patients with epithelial ovarian cancer who were admitted and treated in our hospital and Jilin University Affiliated Hospital, plus 15 blocks of normal ovarian tissue samples were selected. Excised ovarian tissues were tested via immunohistochemistry to test the protein expression of Livin and Beclin1, and their relations with tissue differentiation of ovarian cancer, lymphatic metastasis and clinical staging were analyzed. Results Positive rate of expression of Livin, rare expression in normal ovarian tissues (6.7%), and the highest expression rate in tissues of ovarian cancer (68.6%);Positive rate of expression of Beclin1,the highest expression rate in normal ovarian tissues (93.3%) and the lowest in tissues of ovarian cancer (37.1%). Tissue differentiation of ovarian cancer, lymphatic metastasis and clinical staging were interrelated with the expression degree of Livin and Beclin1 (P<0.05). Conclusion Expression of Livin and Beclin1 in epithelial ovarian cancer and tumor invasion and metastasis plays an important role and may become a new target in the treatment of ovarian cancer.

Expression of livin genes in colon cancer tissues chip and its correlation with bcl-2 genes / 中国现代医生

Objective To discuss expression of livin genes in colon cancer tissues chip and its correlation with Bcl-2 genes. Methods A total of 60 cases of patients with colon cancer, who chose to be treated with surgical operation in General Surgery Department were selected, and applied with immunohistochemical MaxVisionTM two step method to detect the expression of livin and Bcl-2 genesn in tissues chip. 25 cases of colon tissues chip with normal cutting edge were additionally chosen as control group. Results Positive livin genes expression rates in colon cancer tissues chip (63.33%) were much higher than those in colon tissues with normal cutting edge, which appeared statistical differences (24.00%) (χ2=10.93, P<0.01). Livin genes expression in colon cancer tissues chip had no obvious relevance with gen-der, age, tumor size, differentiation degree, lymphatic metastasis and other pathological characteristics (P>0.05), but ob-vious relevance with invasive depth, Duke’s stages, distant metastasis and other pathological characteristics (P<0.05 or P<0.01). Positive Bcl-2 genes expression rates in colon cancer tissues chip (68.33%)were much higher than those in colon tissues with normal cutting edge, which appeared statistical differences (4.00%) (χ2=29.22, P<0.01). Livin genes expres-sion was positively relatedto Bcl-2 genes in expression in colon cancer tissues chip (P<0.01). The invasive depth, Duke’s stages and distant metastasis were included in meta stasis of dinthe Logistic regression model for livin genes expression (P<0.05). Conclusion Livin genes play important roles in the disease process of colon cancer tissues chip and have close relevance with pathologic process of malignant invasion and transfer of tumor. The expression of Livin genes and Bcl-2 genes may play synergetic roles in process of colon cancer.

The expression and clinical significance of Livin and Caspase-9 in duodenal adenocarcinoma / 安徽医科大学学报

The expressions of Livin and Caspase-9 were detected by immunohistochemical technique in 60 duode-nal adenocarcinoma tissues and 30 adjacent normal tissue. The positive expression rate of Livin in duodenal adeno-carcinoma tissues was significantly higher than that of adjacent normal tissue,and the expression of Caspase-9 was lower than that of adjacent normal tissue ( P0.05 ) . Livin and Caspase-9 were associated with occurrence and development of duodenal adeno-carcinoma,and the expression of the above two were negatively correlated. Livin and Caspase-9 may be considered as reliable markers for clinical diagnosis,therapy efficacy and the prognosis of duodenal adenocarcinoma.

Variation of cell proliferous capability and antiapoptosis factors in colon carcinogenesis / 肿瘤研究与临床

Objective To investigate the expression of Ki-67,Survivin,Livin in dysplasia,colon carcinogenesis and para-carcinoma tissues,and to discuss the variation of cell proliferous capability in colon carcinogenesis and antiapoptosis factors.Methods 219 specimens were composed of mild,moderate and severe atypical hyperplasia,well,moderately,poorly differentiated adenocarcinoma,para-carcinoma tissues of 36,34,18,35,27,35 and 34 cases.Detected the expression of Ki-67,Survivin and Livin with tissue microarray and immunohistochemical methods.All data were statistically analyzed by SPSS 17.0.Results In mild,moderate and severe atypical hyperplasia,well,moderately,poorly differentiated adenocarcinoma and para-carcinoma tissues the expression of Ki-67 were (21.56 ± 19.20)％,(37.44 ± 17.41)％,(36.17 ± 17.41)％,(55.29 ± 16.13)％,(44.89 ± 29.67)％,(45.11 ± 29.24)％,(43.94 ± 28.84)％,Survivin were 13.8 ％,44.1％,77.8 ％,85.7 ％,85.1％,91.4 ％,91.1％,and Livin were 2.7 ％,38.2 ％,55.6 ％,100.0 ％,77.8 ％,80.8 ％,79.4 ％.The differences of Ki-67,Survivin and Livin expression in each group were statistically significant (F =6.796,X2 =81.754,X2 =95.200,all P ＜ 0.05).Ki-67 was significantly correlated with expression of Livin (r =0.360,P ＜ 0.05) and no correlated with expression of Survivin (r =0.044,P ＞ 0.05).Conclusion Colonic epithelium from mild atypical hyperplasia to proliferation of well-differentiated adenocarcinoma cells formed a peak,and gradually down to poorly differentiated adenocarcinoma formed a platform.When the colon epithelial cells to become cancerous,the capability of cell proliferation will significantly enhance,apoptosis inhibition will reach the peak and the tumor cell will happen the changes of malignant biological behavior.Tumor microenvironment may promote the cell proliferation in para-carcinoma tissues and the development of colon cancer.Livin may inhibit apoptosis and promot the progression in synergistic mechanism importing with Survivin,which play a role in the development of colonic adenocarcinoma.

Research progress of Livin gene in tumors / 国际外科学杂志

Livin,as a novel member of human inhibitor of apoptosis protein family,is highly expressed in many malignant tumors.Livin plays critical role in apoptosis inhibition,regulating the cell cycle,participating in tumor angiogenesis.Livin is also significant to chemoresistance.The majority of the current data suggests that Livin expression in cancer appears to be associated with unfavorable clinico-pathological parameters,such as disease relapse and shorter patient survival.In recent years,immunotherapy and gene therapy for the targeting of Livin have become a hot research field,which provides new strategy and direction for tumor therapy.

Effect of Livin RNA interference on radiosensitivity of colorectal cancer HT-29 cell xenograft in nude mice / 中华放射医学与防护杂志

Objective To explore the effects of silencing Livin gene by RNA interference mediated by lentiviral vector on colorectal cancer HT-29 cell xenograft growth and sensitivity to radiotherapy in nude mice.Methods BALB/c nude mice models were established by subcutaneously inoculating differently treated HT-29 cells into nude mice and the tumor growth situation of tumors was observed by measuring the volume of tumors and the weight of the nude mice at different time points after cell seeding.Livin expression was detected by RT-PCR and immunohistochemistry,respetively.Apoptosis rate was detected by TUNEL.Normal saline,lentivirus carring unrelated sequences,lentivirus caning Livin shRNA were injected intratumorally.All the nude mice were given 10 Gy of 6 MV X-ray irradiation.The changes of mice weight and the tumor volume were measured at different time points and the weight and tumor growth curves were drawn.Results The inhibition rate of tumor volume was(50.04 ± 0.07)％ and the tumor weight of the RNA interfering group was significantly less than that in experimental group compared to the blank and negative groups(F=4.85,P＜0.05),and the inhibition rate of tumor weight was(50.27 ±0.17)％.Relative Livin mRNA expression level in the RNA interfering experimental group was(17.75 ±0.08)％,and was significantly lower than that of the blank group(67.60 ± 0.05)％ and the negative group(68.54 ± 0.03)％(F=89.97,P＜0.01).Livin protein expression level in the RNA inferring group was also significantly lower[(36.00 ± 3.40)％ versus(85.00 ± 3.15)％,(80.33 ± 3.08)％,F=107.32,P＜0.01].The apoptosis rate in the RNA interfering experimental group was significantly higher than that in the blank and the negative groups[(23.67 ± 2.25)％ versus(5.00 ± 1.50)％,(8.33 ± 1.82)％,F=56.94,P＜0.01].Combined with radiotherapy,the tumor volume at different groups had significant difference(F=10.70,P＜0.01),and RNA interfering group was significantly less than negative group and blank group(F=7.01-9.32,P＜0.01).Conclusions Silencing of Livin gene expression by lentiviral vector-mediated RNA interference could inhibit the growth of colorectal HT-29 cell xenograft and increase the sensitivity of the transplanted tumors to radiotherapy.

Effect of siRNA targeting Livin gene on biological characteristics of human prostate cancer PC3 cells / 中华泌尿外科杂志

Objective To observe the effect of RNAi targeting Livin gene on biology characteristics such as apoptosis and proliferation in human prostate cancer cells.Methods siRNA expression vector targeting Livin gene was constructed and transfected into human prostate cancer cell line PC3.The expressions of Livin mRNA and protein were detected by real-time PCR and Western-blot,cell apoptosis and cell cycle were assayed by flow cytometry,proliferation and colony formation were detected by MTT and colony formation assay,and the tumor growth in vivo was observed in nude mice.Results After transfection,downregulation of Livin mRNA and protein expression in PC3 cells was observed (P＜0.01).Compared with the control group,the proliferation of cancer cells was inhibited significantly (P＜0.01) and the apoptotic ratio was (26.5±3.3) ％ (P＜0.01).The Caspase3 activity increased obviously (P＜0.05),and the experimental group showed a decreased colony formation rate (P＜0.01).The tumor volume of xenografts in nude mouse in experimental and control group was (1.79± 0.07) and (4.40 ± 0.06) cm3 respectively (P ＜ 0.01).Conclusions The siRNA recombinant expression vector targeting Livin gene was constructed and can knockdown the expression of Livin mRNA and protein.It can inhibit PC3 cell proliferation,induce apoptosis and inhibit tumor growth in vivo.

Observation on effect of Livin on VSV-induced apoptosis of A549 cells / 肿瘤研究与临床

Objective To investigate the effect of Livin expression on VSV-induced apoptsis of A549 cells.Methods The expression of Livin of A549 cells was inhibited by RNA interference.VSV-induced apoptosis of A549 cells was observed by Tunel assay.Protein Level of livin was detected by Western blot.Caspase-3 activity was detected by the fluorescence-based quantitative method.Results Livin downregulation VSV-induced apoptosis of A549 cells.Inhibited the expression of Livin of A549 cells had increased Caspase-3 activity.Conclusion The effect of Livin on VSV-induced apoptotic of A549 cells could be increased by RNA interference.

Expressions of apoptosis-regulating proteins in keratoacanthoma and squamous cell carcinoma / 中华皮肤科杂志

ObjectiveTo investigate the difference in cell apoptosis between keratoacanthoma and squamous cell carcinoma.Methods Ninety specimens were obtained from the lesions of 30 patients with regressing keratoacanthoma (rKA) and 30 patients with well-differentiated squamous cell carcinoma (wdSCC) as well as from the normal skin of 30 human controls.Immunohistochemistry was carried out to measure the expressions of B-cell leukemia/lymphoma 2(Bcl-2),cysteine-aspartic acid protease(Caspase)-3,second mitochondria-derived activator of caspase/direct inhibitor of apoptosis-binding protein with low pl (Smac/DIABLO) and inhibitor of apoptosis protein Livin.ResultsThe Bcl-2 protein expression was significantly lower in rKA specimens than in normal control specimens(t =3.1572,P ＜ 0.05).A lower expression of Caspase-3 protein was observed in rKA specimens compared with wdSCC specimens (t =2.1364,P ＜ 0.05).The expression of Smac/DIABLO protein was significantly increased in normal control specimens than in rKA and wdSCC specimens(t =7.6141,9.5666,respectively,both P ＜ 0.05).Livin was absent in normal skin,but highly expressed in rKA and wdSCC specimens,and the difference was significant between the normal skin and lesional (rKA and wdSCC) specimens ( t =4.7913,12.7737,respectively,both P ＜ 0.05).The expression of Livin protein was significantly reduced in rKA specimens than in wdSCC specimens(t =7.9824,P ＜ 0.05).Conclusions Cell apoptosis plays a certain role in the natural regression process of KA.The detection of apoptosis-regulating proteins Caspase-3 and Livin are beneficial for the differentiation between rKA and wdSCC.

Prospective experimental studies on the renal protective effect of ulinastatin after paraquat poisoning / 世界急诊医学杂志（英文）

BACKGROUND: Paraquat (PQ) is an effective herbicide and is widely used in agricultural production, but PQ poisoning is frequently seen in humans with the lung as the target organ. Currently, there are many studies on lung injury after PQ poisoning. But the kidney as the main excretory organ after PQ poisoning is rarely studied and the mechanisms of this poisoning is not very clear. In this study, we observed the expression of caspase-3 and livin protein in rat renal tissue after PQ poisoning as well as the therapeutic effects of ulinastatin. METHODS: Fifty-four Sprague-Dawley (SD) rats were randomly divided into three experimental groups: control group (group A), paraquat poisoning group (group B) and ulinastatin group (group C), with 18 rats in each group. Rats in group B and group C were administered intragastrically with 80 mg/kg PQ, rats in group C were injected peritoneally with 100000 U/kg ulinastatin once a day, while rats in group A were administered intragastrically with the same volume of saline as PQ. At 24, 48, 72 hours after poisoning, the expression of livin in renal tissue was detected by Westen blotting, the expression of caspase-3 was detected by immunohistochemistry, and the rate of renal cell apoptosis was tested by TUNEL detection. The histopathological changes were observed at the same time. RESULTS: Compared to group A, the expression of caspase-3 in the renal tissue of rats in groups B and C increased significantly at any time point. Compared with group B, the expression of caspase-3 in renal tissue of rats in group C decreased. Compared with group A, the expression of livin in renal tissue in rats of groups B and C increased significantly at any time point (P<0.01), especially in group C (P<0.01). TUNEL method showed that the rate of renal cell apoptosis index was higher in group B at corresponding time points than in group A (P<0.01), and was lower in group C at corresponding time points than in group B (P<0.01). CONCLUSION: UTI has a protective effect on the renal tissue of rats after paraquat poisoning through up-regulating the expression of livin and down-regulating the expression of caspase-3, but the regulation path still needs a further research.