诱导型一氧化氮合酶基因转染对人舌癌细胞体内致瘤力的影响

目的研究外源性基因诱导型一氧化氮合酶(iNOS)转染人舌癌细胞系Tca8113后,对肿瘤细胞生物学特性以及裸鼠体内致瘤力的影响。方法采用脂质体转染法将真核表达载体pCMV-iNOS/Tca转染入舌癌细胞系Tca8113,得到高表达iNOS的细胞系pCMV-iNOS/Tca,转染空白质粒的细胞系pCMV-Tca,并以Western blot方法鉴定转染效果。四甲基偶氮唑蓝(MTT)比色绘制生长曲线;流式细胞仪检测细胞周期分布;通过裸鼠成瘤实验,比较转染前后体内成瘤能力的差别。结果与未转染组和转染空白载体组比较,pCMV-iNOS/Tca组的细胞生长速度比Tca组快(P>0.05),细胞周期中G2/M和S期比例增加(P<0.05)。体内成瘤能力试验结果显示,pCMV-Tca、pCMV-iNOS/Tca组和Tca组平均瘤重分别为(1.1±0.24)g、(2.5±0.48)g和(1.3±0.32)g,pCMV-iNOS/Tca组的成瘤力高于pCMV-Tca和Tca组(P<0.01)。结论转染iNOS基因后,Tca8113细胞具有更强的体外增殖能力和体内致瘤能力。

阴道镜联合P16、Ki67免疫组化染色在宫颈癌前病变中的诊断价值

目的:探讨阴道镜下联合应用抑癌基因蛋白16(P16)、原癌基因细胞增殖核抗原(Ki67)免疫组化染色在宫颈癌前病变中的诊断价值。方法:选择2020年1月至2023年6月就诊于清远市妇幼保健院的400例宫颈癌前病变患者作为研究对象,慢性宫颈炎患者400例作为阴性对照,均接受P16、Ki67检测和阴道镜检查,统计检查结果,以组织活检病理学检查结果为准,分析阴道镜联合P16、Ki67免疫组化染色诊断宫颈癌前病变的价值。结果:阴道镜联合P16、Ki67免疫组化染色诊断宫颈癌前病变的灵敏度为100.00%、阴性预测值为100.00%、特异度为97.50%、阳性预测值为97.56%、准确度为98.75%,均高于各方法单一检查,差异均具有统计学意义(P <0.05)。结论:阴道镜、P16、Ki67检查均是鉴别宫颈癌前病变的有效手段,三者联合检查可提高宫颈癌前病变的检出效果。

联合检测SOX4、P16、Ki67在宫颈上皮内瘤变中的表达及临床意义

目的探究SOX4、P16、Ki67在宫颈上皮内瘤变中的表达及临床意义。方法选取我院2022年6月至2023年5月期间病理科确诊为宫颈上皮内瘤变患者32例为观察组,依据宫颈上皮内瘤变组织病变程度分为两组,其中低级别宫颈上皮内瘤变组织为低级别组(n=16),高级别宫颈上皮内瘤变组织为高级别组(n=16),另选取同期收入10例子宫肌瘤至我院实施全子宫切除术正常宫颈组织患者为对照组,并对上述患者均开展SOX4、P16、Ki67检测,分析表达情况。结果观察组中SOX4、P16、Ki67阳性率分别为50.00%、68.75%、68.75%,对照组均为0例(0.00%),观察组整体阳性率高于对照组,差异有统计学意义(P<0.05);低级别宫颈上皮内瘤变患者中SOX4、P16、Ki67阳性分别为7例(43.75%)、11例(68.75%)、11例(68.75%),高级别宫颈上皮内瘤变患者中分别为10例(62.50%)、13例(81.25%)、10例(62.50%),两组对比差异无统计学意义(P>0.05);SOX4、P16、Ki67阳性表达与宫颈上皮内瘤变呈正相关,P<0.05。结论SOX4、P16、Ki67表达与宫颈上皮内瘤变相关联,参与疾病发生、发展过程,在后续可作为判断宫颈癌疾病发生、发展的生物标志物。

PCNA,C-erbB-2,Cyclin D1及MMP2的表达与胃癌预后因子的相关性分析

为探讨PCNA ,C erbB 2 ,CyclinD1及MMP2等标志物在胃癌中的表达及其与胃癌预后的相关性。应用免疫组织化学染色法检测 61例胃癌组织中PCNA ,C erbB 2 ,CyclinD1和MMP2的表达 ,并将其与患者的性别、年龄、肿瘤大小、组织分化程度、浸润深度及转移淋巴结数目进行相关性分析。结果示PCNA ,C erbB 2 ,CyclinD 1及MMP2均与肿瘤浸润深度和淋巴结转移数目呈正相关。PCNA和CyclinD 1的表达还分别与肿瘤大小和组织分化程度呈正相关 ,而C erbB 2及MMP2与肿瘤大小及组织分化程度无关。提示肿瘤浸润深度和淋巴结转移数目是影响胃癌预后的最重要的独立危险因子 ;胃癌愈到晚期 ,PCNA ,C erbB 2 ,CyclinD1及MMP2等肿瘤标志物表达愈强 ,从而加速了胃癌的生长与转移。

Ki-67联合p16检测对HPV持续高危阳性患者子宫颈高级别鳞状上皮内病变的诊断价值

目的:分析原癌基因细胞增殖核抗原(Ki-67)联合细胞周期蛋白激酶的抑制因子(p16)检测对人乳头瘤病毒(HPV)持续高危阳性患者子宫颈高级别鳞状上皮内病变的诊断价值。方法:选取新余市人民医院2017年12月—2020年11月收治的慢性子宫颈炎患者134例、子宫颈低级别鳞状上皮内病变131例、子宫颈高级别鳞状上皮内病变129例、HPV持续高危阳性患者118例,分别记为对照1组、对照2组、对照3组、研究组,四组均取病灶组织采用免疫组化二步法检测Ki-67、p16蛋白表达,比较四组病灶组织Ki-67、p16蛋白表达情况及阳性率;比较研究组不同临床病理特征患者病变组织Ki-67、p16蛋白表达阳性率;采用列联表法分析在各组子宫颈高级别鳞状上皮内病变与病灶组织Ki-67、p16蛋白表达的关系,采用受试者操作特征(ROC)曲线分析病变组织标本中Ki-67、p16及联合诊断子宫颈高级别鳞状上皮内病变的价值。结果:四组病灶组织Ki-67、p16蛋白表达分布比较差异均有统计学意义(Z=29.852,P<0.001;Z=25.856,P<0.001),研究组病灶组织Ki-67、p16蛋白表达阳性率均高于其余三组(P<0.01),且对照2组、对照3组病灶组织Ki-67、p16蛋白表达阳性率均高于对照1组(P<0.01),研究组子宫颈高级别鳞状上皮内病变与病灶组织Ki-67、p16蛋白表达均呈正相关(C=0.745、0.641,P<0.01),HPV持续高危阳性患者Ki-67、p16及联合诊断子宫颈高级别鳞状上皮内病变的敏感度分别为65.40%、69.20%、78.90%,特异度分别为71.70%、73.60%、73.60%,AUC分别为0.718、0.729、0.816,联合诊断的AUC值高于Ki-67、p16单一诊断(Z=6.854,P=0.000;Z=6.741,P=0.000)。结论:子宫颈高级别鳞状上皮内病变与HPV持续高危阳性患者病灶组织Ki-67、p16蛋白表达均呈正相关,Ki-67联合p16检测的诊断价值较高。

TTYH2,a human homologue of the Drosophila melanogaster gene tweety,is up-regulated in colon carcinoma and involved in cell proliferation and cell aggregation

AIM: To investigate the expression patterns of TTYH2 in the human colon cancer and colon cancer cell lines and to evaluate the inhibitory effect of small interfering RNA (siRNA) on the expression of TTYH2 in colon cancer cell lines.METHODS: We investigated the expression patterns of TTYH2 in colon cancer, adjacent non-tumorous colon mucosa, and cancer cell lines (DLD-1, caco-2, and Lovo) by RT-PCR. Furthermore, a siRNA plasmid expression vector against TTYH2 was constructed and transfected into DLD-1 and Caco-2 with LipofectamineTM 2000. The down regulation of TTYH2 expression was detected by RT-PCR and the role of siRNA in inducing cell proliferation and cell aggregation was evaluated by MTT and aggregation assay.RESULTS: TTYH2 gene expression in colon cancer tissue was significantly up-regulated compared with normal colonic mucosa (1.23 ± 0.404 vs 0.655 ± 0.373, P = 0.0103). Colon cancer derived cell lines including DLD-1, Caco-2, and Lovo also expressed high levels of TTYH2. In contrast, transfection with siRNA-TTYH2 signifi cantly inhibited both proliferation and scattering of these cancer cell lines.CONCLUSION: The present work demonstrates, for the fi rst time, that the TTYH2 gene expression is signifi cantly up-regulated in colon cancer. The TTYH2 gene may play an important role in regulating both proliferating andmetastatic potentials of colorectal cancer.

PlexinA1 expression in gastric carcinoma and its relationship with tumor angiogenesis and proliferation

AIM: To explore the expression of PlexinA1 in gastric carcinoma and its relationship with tumor angiogenesis and proliferation. METHODS: PlexinA1 mRNA and protein expressions of Semaphorin6D were measured using semi-quantity reverse transcription PCR and Western blotting in 20 cases of gastric carcinoma and corresponding normal gastric mucosa. PlexinA1, Ki-67 expression and microvessel density (MVD) were detected by immunohistochemistry in 50 cases of gastric carcinoma and 20 cases of normal gastric mucosa. RESULTS: The mRNA and protein expressions of PlexinA1 in gastric carcinoma were significantly higher than that in normal gastric mucosa (0.71 ± 0.37 vs 0.60 ± 0.25, P = 0.0299 < 0.05, and 0.47 ± 0.16 vs 0.21 ± 0.08, P = 0.0000 < 0.01), and MVD within tumor tissues increased significantly with PlexinA1 mRNA expression (r =0.8736, P < 0.01) and PlexinA1 protein expression (r = 0.7286, P < 0.01), and MVD of the PlexinA1 positive staining group (25.25 ± 3.93) was significantly higher than that of the negative group (19.56 ± 1.75), (P < 0.01). Proliferation index of tumor cells within tumor tissues were positively correlated with PlexinA1 mRNA expression (r = 0.5420, P = 0.014 < 0.01) and PlexinA1 protein expression (r = 0.5024, P = 0.024 < 0.05). The proliferation index of the PlexinA1 positive staining group (567.69 ± 125.61) was signifi cantly higher than that of the negative group (369.58 ± 116.88), (P < 0.01). CONCLUSION: PlexinA1 may play an important role in the occurrence and development of gastric carcinoma, and be related to tumor angiogenesis and proliferation.

S100A6 gene have a positive influence on the growth and proliferation of gastric cancer cell MKN45

Objective： S100A6 （a.k.a., calcyclin） is over-expressed in several human tumors, including gastric carcinoma, human melanoma, pancreatic carcinoma, squamous cell carcinoma, malignant fibrous histiocytoma （MFH）, and carcinomas of the thyroid, breast, and colon. However, little is known about the role S100A6 plays in gastric adenocarcinoma. In the present study, we intended to investigate the influence of S100A6 on the growth, proliferation, apoptosis, invasion and cell cycle of the gastric cancer cell MKN45. Methods： As an important member of S100 family, S100A6 cDNAwas subcloned into a constitutive vector pcDNA3.1 followed by transfection in gastric cancer cell line MKN45 by using liposome. Then stable transfectants were selected and appraised. The apoptosis and cell cycles of these clones were analyzed by using flow cytometric assay. The growth and proliferation were analyzed by cell growth curves and colony-forming assay respectively. The S100A6 stable expression clones （MKN-S100A6） were detected and compared with their control groups respectively. Results： MKN- S100A6 grew faster than MKN45 and MKN-PC （MKN45 transfected with pcDNA3.1 vector）. The cell counts of MKN-SI00A6 in the fifth, sixth and seventh days were significantly more than those of control groups （P 〈 0.05）. Cell cycle analysis showed that proportions of MKN-S100A6 in G0-G1 and G2-M were different significantly with those of its control groups respectively （P 〈 0.05）. The apoptosis rate of MKN-S100A6 was significantly lower than those of control groups （P 〈 0.05）. Results of colony-forming assay showed that the colon formation rate of MKN-S100A6 was higher than those of control groups （P 〈 0.05）. Conclusion： S100A6 can promote the growth and proliferation of gastric cancer cells. It can help tumor cell maintain malignant phenotype. In gastric cancer, S100A6 could be thought as a tumor-enhancing gene in some distance, but its role could be complicated.

Inhibitory effect of adeno-associated virus-mediated gene transfer of human endostatin on hepatocellular carcinoma

AIM: To investigate the effect of adeno-associated virusmediated gene transfer of human endostatin on the growth of hepatocellular carcinoma (HCC).METHODS: HCC cell line Hep3B was infected with recombinantadeno-associated virus containing human endostatin gene (rAAV2-hEndo). The results of transfection were detected by RT-PCR and SDS-PAGE assay. MTT assay was used to observe the effects of supernatant of transfected cells on ECV304 cell proliferation. An animal model of HCC was established by injecting Hep3B cells subcutaneously into the back of nude mice. Intratumoral injection of rAAV2hEndo, empty virus and phosphate-buffered saline were given sequentially. Serum endostatin was determined byELISA, the inhibitory effect of endostatin on the growth of xenograft was assessed in 3 wk.RESULTS: The results of RT-PCR and SDS-PAGE assay confirmed that rAAV2-hEndo successfully transfected Hep3B cells, and endostatin was secreted from Hep3B cells to medium. The supernatant of transfected cells markedly inhibited the proliferation of ECV304 cells (P＜0.01). Intratumoral injection of rAAV2-hEndo (2×1010v.g.) led to a sustained serum endostatin level ofapproximately (86.71±5.19) ng/mL. The tumor volumeand microvessel density were less in rAAV2-hEndo group than in control groups (P＜0.01).CONCLUSION: Human endostatin can be stably expressed by adeno-associated virus-mediated gene transfer and effectively inhibit the growth of HCC.

Cloning and characterization of human IC53-2,a novel CDK5 activator binding protein

We have identified IC53-2, a human homologue of the rat C53 gene from a human placenta cDNA library (GeneBank Accession No. AF217982). IC53-2 can bind to the CDK5 activator p35 by in vitro association assay. IC53-2 is mapped to human chromosome 17q21.31. The IC53-2 transcript is highly expressed in kidney, liver, skeletal muscle and placenta. It is abundantly expressed in SMMC-7721, C-33A, 3AO, A431and MCF-7 cancer cell lines by RT-PCR assay. Stable transfection of IC53-2 cDNA into the hepatocellularcarcinoma SMMC-7721 cell remarkably stimulates its growth in vitro. The above results indicate thatIC53-2 is a novel human gene, which may be involved in the regulation of cell proliferation.

The effects of interfering in COX-2 gene expression on malignant proliferation of the human lung adenocarcinoma A2 cell in vitro

Objective:We explored the interfering effect of COX-2 gene expression and the influence on the malignant proliferation of A2 cells by RNAi after quenching COX-2 in vitro. Methods:COX-2 was selected as the subject. Three COX-2 siRNA expression vectors with human U6 promoter were constructed and three vectors and the vacant vector (pEGFP) were transfected into A2 cells with lipofectamine respectively. The cell strains transfected were constructed. The change of COX-2 expression levels was examined by Western blot and RT-PCR. The effects on the proliferation of A2 cells after silencing COX-2 were studied by cell growth curve and clonogenic assay in vitro. Results: The three siRNA and U6 promoter cloned into pEGFP plasmid were validated by PCR, restriction endonucleases identification, DNA sequencing and BLAST alignment. The cell strains transfected were named as A2-3, A2-7, A2-10 and A2-p respectively. Green fluorescence was seen in A2-p cells and not in A2-3, A2-7 and A2-10 cells in 24, 48 and 72 h after transfected. The results of RT-PCR and Western blot showed the three siRNA expression vectors produced effects and the expression of COX-2 was inhibited in different extent. In contrast to A2 cells, the levels of COX-2 mRNA of A2-3, A2-7 and A2-10 cells reduced 15.6%, 20.4% and 64.2% respectively; the levels of COX-2 protein of A2-3, A2-7 and A2-10 cells reduced 23.7%, 36.7% and 60.2% respectively. The results of cell growth curve and clonogenic assay showed the growth of A2-10 cell slowed and the colonial formation rate reduced but the growth of A2-3 and A2-7 cells had not obvious changes in contrast to the controls (A2 and A2-p). Conclusion: Silencing the COX-2 gene in vitro by RNAi technique can significantly inhibit the malignant proliferation of A2 cells.

Effect of hammerhead ribozyme that specifically cleaves c-myc mRNA on proliferation of vascular smooth muscle cells

Ojective: To investigate the effect of hammerhead ribozyme that specifically cleaves c-myc mRNA on the proliferation of vascular smooth muscle cells (VSMCs ). Methods: Based on the computer analysis of the secondary structure of c-myc mRNA, nt 2029 in rat c-myc oncogene was selected as a cleaving site for hammerhead ribozyme and the ribozyme was designed. With automatic DNA synthesizer, the two complementary DNA strands of the ribozyme were synthesized. The ribozyme gene was cloned into pGEM3Zf ( + ) vector and subcloned into eukaryotic expression pcD-NA3 vector. The recombinant pcDNA-Rz was transfected into the cultured rat VSMCs by lipofectAMINE mediated DNA transfection protocol and individual cell clones were selected by G418. Results: The sequence of ribozyme gene inserted in pGEMSZf ( + ) vector was proved to be perfectly correct. In VSMCs transfected with recombinant pcDNA-Rz, flow cytometry analysis showed that the S phase and G2/M fractions were decreased significantly and cell proliferation stagnated in the G0/G1 phase. Conclusion: The results suggest that hammerhead ribozyme that specifically cleaves c-myc mRNA can significantly inhibit the proliferation of VSMCs.

Downregulation of wild-type p53 protein by HER-2/neu mediated PI3K pathway activation in human breast cancer cells: its effect on cell proliferation and implication for therapy

Overexpression and activation of HER-2/neu (also known as c-erbB-2), a proto-oncogene, was found in about 30% of human breast cancers, promoting cancer growth and making cancer cells resistant to chemo- and radio-therapy. Wild-type p53 is crucial in regulating cell growth and apoptosis and is found to be mutated or deleted in 60-70% of human cancers. And some cancers with a wild-type p53 do not have normal p53 function, suggesting that it is implicated in a complex process regulated by many factors. In the present study, we showed that the overexpression of HER-2/neu could decrease the amount of wild-type p53 protein via activating PI3K pathway, as well as inducing MDM2 nuclear translocation in MCF7 human breast cancer cells. Blockage of PI3K pathway with its specific inhibitor LY294002 caused Gl-S phase arrest, decreased cell growth rate and increased chemo- and radio-therapeutic sensitivity in MCF7 cells expressing wild-type p53. However, it did not increase the sensitivity to adriamycin in MDA-MB-453 breast cancer cells containing mutant p53. Our study indicates that blocking PI3K pathway activation mediated by HER-2/neu overexpression may be useful in the treatment of breast tumors with HER-2/neu overexpression and wild-type p53.

Decreasing activity of NF-κB inhibits the proliferation of A549 cells via the down-regulation of cyclin D1 expression

Objective: To investigate the effect of decreasing activity of NF-κB on proliferation of A549 cell line and the pos-sible molecular mechanism. Methods: The recombinant plasmid PCDNA3.1(+)/IκBα expressing IκBα was constructed. The recombinant plasmid was then transfected to A549 cell. The activity of NF-κB, cell proliferation, and cyclin D1 expression were observed. Results: Our results showed that transfecting PCDNA3.1(+)/IκBα inhibited activity of NF-κB in A549 cells, and decreasing activity of NF-κB inhibited proliferation of A549 cells. Decreasing activity of NF-κB was accompanied with down-regulation of cyclin D1 expression. Conclusion: Decreasing activity of NF-κB inhibited proliferation of A549 cells, and the molecular mechanism of the inhibition effect may be down-regulation of cyclin D1 expression.

Influence of Deleted in Colorectal Carcinoma Gene on Proliferation of Ovarian Cancer Cell Line SKOV-3 In Vivo and In Vitro

Objective To elucidate the effects of the deleted in colorectal carcinoma(DCC) gene on proliferation of ovarian cancer cell line SKOV-3.Method An exogenous recombinant eukaryotic expression vector pcDNA3.1(+)-DCC,containing human DCC cDNA coding sequences,was constructed and transfected into SKOV-3 cells(SKOV-3/DCC).The pcDNA3.1(+) transfected cells(SKOV-3/Neo) and SKOV-3 cells were used as the positive and negative controls,respectively.Expressions of DCC mRNA and protein were analyzed by RT-PCR and immunocytochemical analysis,respectively.Cell growth was detected by soft agar colony formation assay and MTT assay.Flow cytometry and transmission electron microscopy were used to assess the effects of DCC on cell cycle distribution and ultrastructure,respectively.BALB/c mice were used to evaluate the effects of DCC on tumorigenicity in vivo.Results RT-PCR and immunocytochemical analysis revealed the exogenous DCC gene was successfully transfected into SKOV-3 cell lines and obtained permanent expression.The half maximal inhibitory concentration(IC50) of SKOV-3/DCC cells was significantly lower than that of SKOV-3 or SKOV-3/Neo cells(all P<0.05).DCC expression caused SKOV-3 cells to be arrested in G1 phase(78.0%),and electron microscopic analysis showed SKOV-3/DCC cells displayed typical morphological changes of apoptosis.Two mice xenografted with SKOV-3/DCC cells showed no tumor tumorigenecity.The tumor volume of BALB/c mice bearing SKOV-3/DCC cells(3.403 mm3) was smaller than that of SKOV-3 cells(9.206 mm3).Conclusion DCC gene may play an important role in suppressing the growth of SKOV-3 cell line and inducing apoptosis.

血管紧张素Ⅱ诱导新生鼠心肌细胞凋亡及c-fos、PCNA表达意义

目的研究血管紧张素Ⅱ(angiotensinⅡ AngⅡ)对新生鼠原代培养心肌细胞凋亡及c-fos、PCNA蛋白表达变化的影响。方法采用体外心肌细胞培养技术,培养Wistar新生鼠原代心肌细胞,将培养96h的心肌细胞随机分为两组,对照组:无血清培养2、6、12、24h。AngⅡ组:以10-7mol/LAngⅡ刺激2、6、12、24h,用TUNEL(terminal deoxynucleotidyl transferse-mediated dUTP nick-end labeling assay)方法检测凋亡细胞数,免疫组化方法观察c-fos、PCNA(proliterating cell nuclear angiten)蛋白染色。结果随着时间的延长AngⅡ组TUNEL染色凋亡心肌细胞数量与对照组相比显著增高;同时伴有早期c-fos、PCNA蛋白表达增加。结论AngⅡ可以诱导乳鼠心肌细胞凋亡,AngⅡ诱导心肌细胞凋亡早期可见c-fos、PCNA表达增强。

Effect of Lentivirus-induced shRNA Silencing CXCR4 Gene on Proliferation and Apoptosis in Human Esophageal Carcinoma Cell Line Eca109

OBJECTIVE To discuss the application of the slow virus-induced short-hairpin RNA （vshRNA） to silence the expression of CXCR4 in EsCa cell lines Eca109, and observe the effect of silencing CXCR4 on the proliferation and apoptosis of Eca109 cells in vitro. METHODS The expression plasmid of vshRNA targeting CXCR4 was constructed, with a concurrent construction of negative vshRNA expression plasmid, and without targeting any known mRNA. Real-time quantitative PCR and Western blot assay were used to determine the change of CXCR4 expression in the post-transfected EsCa cell Eca109, and MTT assay was conducted to detect the change of proliferation in EsCa Eca109 cell after silencing the CXCR4. The .ow cytometry was used to detect the change of the cell cycle and apoptosis in the post-silenced EsCa Eca109 cell in di. erent groups. RESULTS The transfection rate was respectively （87.3 ± 1.2）% and （90.1 ± 1.4）% in the CXCR4- RNAi-LV （silent group） and NC-GFP-RNAi-LV （negative control group） cellular plasmids. The vshRNA interference resulted in a down-regulation of the CXCR4 gene mRNA and protein expressions in Eca109 cells. CXCL12 promoted the proliferation of EsCa cell lines Eca109. The speed of EsCa cell proliferation became slower in the silencing group than in the normal control （also the control） and the negative control groups （P 〈 0.05）. However, there was no significant difference in comparison of the proliferation speeds between the negative control and the normal control groups （P 〉 0.05）. In the silencing group, the proportion of the cells in phase G0/G1, phase S and phase G2/M was respectively （69.9 ± 5.0）%, （17.1 ± 2.5）% and （13.0 ± 7.4）%, and the apoptotic rate achieved （7.27 ± 0.50）%. In the normal control group, the proportion of the cells in phase G0/G1, S and G2/M was respectively （55.9 ± 4.6）%, （30.2 ± 3.9）% and （13.8 ± 1.4）%, and the apoptotic rate was （3.30 ± 0.70）%. In the negative control group, the proportion of cells in phase G0/G1, S and G2/M was respectively （52.7 ± 7.8）%, （25.3 ± 2.3）% and （21.9 ± 7.4）%, with an apoptotic rate of （4.03 ± 1.37）%. Compared with the normal control and negative control groups, there was an apparent growth of cells in the phase G0/G1 （P 〈 0.05）, and a greatly increased number of cells in phase S （P 〈 0.05） in the silencing group. There was no signi. cant di.erence in comparison of those between the normal control and negative control groups （P 〉 0.05）. The apoptotic rate was obviously higher in the cells of the silencing group than in the normal control and the negative control groups （P 〈 0.05）. There was no signi. cant di.erence in comparison of the apoptotic rate between the normal control and the negative control groups （P 〉 0.05）. CONCLUSION CXCR4-vshRNA can specifically and effectively inhibit CXCR4 expression of Eca109 cells. CXCR4-vshRNA can inhibit the proliferation and enhance the apoptosis rate of Eca109 cells through intervening the expression of CXCR4, suggesting that CXCL12/CXCR4 might have an important role in the progression of Escc Thisslow virus-induced shRNA can effectively silence the expression of CXCR4 gene in the EsCa cells; block up the biological e.ect of CXCL12/CXCR4 axle; and e.ectively inhibit the potency of proliferation in the EsCa cell line Eca109, thus advancing apoptosis. It suggests that the CXCL12/CXCR4 plays an important role in the progression of EsCa.

Demethylation of FANCF gene may be a potential treatment through inhibiting the proliferation of cervical cancer

Objective: The aim of the study was to explore the effect of demethylating agent 5-Aza-2'-deoxycytidine (5-ADC) on expression of Fanconi anemia complementation group F (FANCF) gene and the proliferation of cervical cancer cells, to observe cell's sensitivity to chemotherapeutic drug taxol, and to explore the antitumor effect of 5-ADC as well as the new treatment of cervical cancer. Methods: Cervical cancer cell lines SiHa (FANCF gene full-methylated) and Hela (unmethylated) were treated with 5-ADC. We used the methylation-specific PCR (MSP), reverse transcription-polymerase chain reaction (RT-PCR) and Western blot to detect the FANCF methylation, mRNA and protein respectively. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to detect the proliferation of cells. The cytotoxicity of taxol was measured by flow cytometer. The nude mice bearing SiHa was used to observe the effect of 5-ADC in vivo. Results: Inhibition of DNA promoter methylation by 5-ADC reactivated the expression of FANCF mRNA and protein in SiHa cells, consistent with decreased growth speed and increased taxol resistance. These results were proven in experiments in vivo. Conclusion: The 5-ADC probably become a potential treatment drug through inhibiting the proliferation of cervical cancer cells in taxol-resistant patients.

Polyphyllin I inhibits proliferation and metastasis of ovarian cancer cell line HO-8910PM invitro

OBJECTIVE： To study the anticancer mechanism of polyphyllin I （PPI）, a Traditional Chinese Medicine, on the ovarian cancer cell line HO-8910PM in vitro. METHODS： Transwell chamber invasive assays were used to investigate the inhibitory capacity of PPI on HO-8910PM metastasis. Gene expression profiling chips was used to screen differentially ex- pressed genes between experiment group and con- trol group. Reverse transcription PCR and Western blotting were used to determine mRNA and pro- tein levels. RESULTS： With increasing PPI concentration, the metastatic capacity of cells decreased, with signifi- cance differences between the experimental and control groups （P〈0.01） as well as between two concentration groups. Gene expression profiling identified 123 differentially expressed genes, of which 70 were downregulated and 53 were upregu- lated. The genes were involved in multiple signal transduction pathways, including apoptosis, prolif- eration and metastasis. Real-time PCR （RT-PCR） showed that differential genes PIK3C2B, Caspase 9and WntSA were downregulated with increasing PPI, showing an evident dose-effect relationship. The c-Jun was an exception. As the PPI dosage in- creased and the exposure time was extended, c-Jun relative expression showed an upward trend. There were significant differences between the ex- periment and control （P〈0.05）. Western blot analy- ses showed that PPI treatment decreased levels of Caspase 9, WntSA and PIK3C2B and increased acti- vated Caspase 9,c-Jun and p-c-Jun expression levels. CONCLUSION＂ PPI has strong antitumor and anti transfer activity. It can activate c-Jun expression and the JNK signaling pathway, elicit cell apoptosis via the mitochondrial-mediated Caspase activation pathway, and finally inhibit tumor growth and mi- gration in vitro. The downregulation of PIK3C2B and Wnt5A jointly inhibit the proliferation and me- tastasis of HO-8910PM. PPI may be a novel treat- ment for ovarian cancer.