Advances of homeobox gene MEOX1 in tumors / 国际肿瘤学杂志

As a homeobox transcription factor,MEOX1 can regulate the target genes by binding the specific DNA sequence.MEOX1 not only plays essential roles in cell proliferation,migration and differentiation,but also participates in the formation of skeleton,muscle and blood vessel during embryonic development.Recent studies demonstrate that MEOX1 is over-expressed in breast cancer,lung cancer,ovarian cancer and prostate cancer tissues,which is closely associated with lymph node metastasis and poor prognosis in patients with cancer.Furthermore,MEOX1 can regulate the proliferation and migration of cancer cells,which suggests that it plays an important role in the occurrence and development of tumors.

Roles of HOXC gene in tumors / 国际肿瘤学杂志

The protein encoded by the HOXC gene regulates multiple processes during embryonic development,including cell growth,differentiation,apoptosis,migration and angiogenesis.The HOXC gene is also involved in the occurrence and development of various human solid tumors,such as colorectal cancer,gastric cancer,liver cancer,breast cancer,endometrial cancer,etc.In addition,the HOXC gene is closely related to the clinical stage and distant metastasis of the tumor.Thus,the HOXC gene can be used as a biological predictor of tumor metastasis and prognosis.

Effects of 5-Aza-2'-deoxycytidine on proliferation of human gastric cancer BCG-823 cells and the expression of HOXA5 gene / 中国医师杂志

Objective To explore the effects of 5-Aza-2'-deoxycytidine (5-Aza-CdR) on the proliferation of BCG-823 human gastric cancer cells and the expression of HOXA5.Methods The methylation status of the promoter of HOXA5 was measured by methylmion specific PCR (MSP).The BCG-823 cells were treated with different concentrations of 5-Aza-CdR,and then the changes of expression and methylation status of HOXA5 gene were detected by quantitative real-time polymerase chain reaction (QRT-PCR),Western-blot,and MSP.Cells proliferation was assessed by methyl thiazolyl tetrazolium (MTT) assay.Results (1) Different methylation status of HOXA5 gene promoter was detected in BCG-823 cells.The mathylation rate of HOXA5 gene promoter were reduced after treatment with 5-Aza-CdR,and also were negatively related to the concentration of 5-Aza-CdR (F =438.307,P ＜ 0.01).(2) Compared to the control groups,the expressions of HOXA5 mRNA and protein were increased after treatment with 5-Aza-CdR,with statistical significance (P ＜ 0.05).(3) The proliferation rate of BCG-823 cells was significantly inhibited (P ＜0.05).Conclusions The methylation status of HOXA5 promoter was detected in BCG-823 cells.5-AzaCdR is able to inhibit BCG-823 cells growth in vitro,which might be related to the expression of HOXA5.It may be a new way to treat gastric cancer.

Expression and role of HOXC gene in tumor / 国际肿瘤学杂志

As a member of homeobox gene family,HOXC is expressed in many organs and can regulate gene expression,cell differentiation and morphogenesis.Abnormality of its function is closely related to the prognosis of leukemia,breast cancer,renal cell carcinoma,prostate cancer and so on.

The construction of human prostate specific homeobox gene NKX3.1 eukaryotic expression vector and preliminary study on the biological function of NKX3.1 / 中国医师杂志

Objective To investigate the basic biological functions of NKX3.1 with construction of a series of eukaryotic expression vector of NKX3.1 to study its effects on cell cycle and apoptosis.Methods The full-length reading frames (ORF) of NKX3.1 was amplified from normal human prostate tissue with polymerase chain reaction (PCR),the ORF was cloned into T vector,and then NKX3.1 was cloned into eukaryotic expression vector pcDNA3.1 (+) and pcDNA3.1 (-) with this T vector as template.NKX3.1 in pEGFP C1 was transfected into PC3 cell line,and C1 pEFP was taken as control to observe the localization of NKX3.1 in cells.At the same time,NKX3.1 in pcDNA3.1 and pcDNA3.1 were transfected into PC3 cells and the cells were collected after 24 ～48 hours to detect cell cycle and apoptosis by flow cytometry.Results The eukaryotic expression vector of NKX3.1 was constructed successfully.Fluorescent protein was ocated in the nucleus after transfection of NKX3.1 in pEGFP C1,but pEGP C1 was both expressed in nucleus and cytoplasm.There was significant effect on the early apoptosis (P＜0.01),but no effect on the cell cycle (P ＞ 0.05) after transfection of NKX3.1 compared to PC3 cell-transfected with empty vector of pcDNA3.1.Conclusions The result data suggest that NKX3.1 be nuclear protein and have effect on the early apoptosis of PC3 cell line based on the construction of eukaryotic expression vector.

Expression and significance of Six1 in cervical cancer / 天津医药

Objective To investigate the clinical significance of protein expression of Six1 in cervical cancer. Meth?ods The immunohistochemical (IHC) staining was applied to detect the expression of Six1 protein in normal cervical tis?sues (n=32), cervical intraepithelial neoplasia (CIN) tissues (n=49) and cervical cancer tissues (n=123). The localization of Six1 protein was detected in vitro cultured HeLa cells using immunofluorescence (IF) staining. Results The positive rate of Six1was significantly higher in cervical cancer (72.3%) than that of CIN tissues (28.6%) and normal cervical tissues (15.6%,χ2=13.118 and 10.058 respectively, P<0.01). There were significance differences in expression levels of Six1 protein be?tween different tumor sizes and metastasis of cervical cancer (P < 0.01). The Six1 protein showed positive signals in cyto?plasm and nucleoli in HeLa cells. Conclusion Six1 expression is associated with cervical cancer, which may be a potential biomarker for invasion and metastasis of cervical cancer.

Current research of role of Hox genes in pathogenesis of equinus deformity / 天津医药

Congenital clubfoot(CCF), which is also known as equines deformity, is a common congenital malformation that affect children′s life quality. However, its cause is still to be elucidated. Currently, polygenetic and environmental fac?tors are both believed to play important roles in CCF pathogenesis. Several genes including HOX, PITX1, NAT2, P63, DTDS and COL9A were shown to contribute to congenital clubfoot, but which is the most critical gene remains unclear. Several re?ports have revealed that Hox genes are closely related to the cause of CCF. Hox genes are regulators of body morphogenesis, and its mutation result in limbs and trunk deformity in human. Here, we systematically reviewed the latest literature that stud?ied the role of Hox genes in pathogenesis of Congenital clubfoot, with the prospect of laying a foundation for its future clinic treatment.

Effects of ZHX3 gene silence on the expression of osteoblast-related factors in BMSCs / 天津医药

Objective To investigate the effects of zinc fingers and homeoboxes 3 (ZHX3) silence on expressions of smad3, smad4 and RUNX2 in bone marrow mesenchymal stem cells (BMSCs). Methods ZHX3 low expression vector (ZHX3 silent group) was constructed and was transfected to rat BMSCs. Empty vector was transfected into BMSCs and was used as vehicle control group, and wild type BMSCs was used as the control group. The cell transfection rate was measured under a fluorescence microscope, and then the successful transfection was identified. The immunocytochemistry and immu?noblotting methods were used to detect the expression levels of smad3, smad4 and RUNX2. Results (1) Cells with BMSCs phenotype can be obtained by recovery culturing. (2) After transfection, the green fluorescent protein was found in ZHX3 si?lence group and vehicle control group. Blank control group showed no significant fluorescence. The expression level of ZHX 3 was significantly lower in ZHX3 silence group than that of vehicle control group. (3) Results of immunofluorescence asssay showed that the positive expressions of smad3 and smad4 were located in nucleus and cytoplasm, the positive expression of RUNX2 was mainly located in nucleus. Positive cells were observed in three groups. There was no significant difference in fluorescence intensity between the control group and the vehicle control group, but the fluorescence intensity was significant?ly lower in ZHX3 gene silence group than that of two control groups. (4) There were no significant differences in expressions of smad3, smad4 and RUNX2 betweem control group and the vehicle control group, but they were significantly higher than those of ZHX3 silence group(P < 0.05). Conclusion ZHX3 gene silence can delay vitro osteogenesis of BMSCs, which may play a role by the down-regulated expression levels of smad3, smad4 and RUNX2.

Selection and functional identification of siRNA targeting of HOXA7 / 肿瘤研究与临床

Objective To design and screen small interefere RNA (siRNA) targeting of HOXA7,and to investigate the effect of the siRNA on human lung cancer LETP-a-2 cells proliferation and apoptosis in vitro.Methods Three pairs of siRNA targeting of HOXA7 and one pair of siRNA for negative control were transfected respectively into LETP-a-2 cells through cationic liposome.The mRNA and proteion expression levels of HOXA7 were observed by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis.The effect of HOXA7 siRNA on growth and apoptosis of LETP-a-2 cells were measured by MTT and flow cytometry.Results All the three pairs of siRNA could inhibit HOXA7 expression effectively,among which siRNA2 got the best effects,the silence rates were (57.344±4.743) ％ on mRNA level and (52.219±0.550) ％ on protein leval.The proliferation was inhibited and the apoptosis was promoted by the siRNA targeting HOXA7 in LETP-a-2 cells,among which siRNA2 got the favourite results,the inhibitory rate was (48.144±4.992) ％ and the apoptosis rate was (26.613±0.612) ％.Conclusion The siRNA2 targeting of HOXA7 enrolls in promoting apoptosis and inhibiting grows of LETP-a-2 cells,indicating that manipulation of HOXA7 expression may be a potential therapeutic strategy for cancer.

The Mechanism of White and Brown Adipocyte Differentiation

Obesity gives vent to many diseases such as type 2 diabetes, hypertension, and hyperlipidemia, being considered as the main causes of mortality and morbidity worldwide. The pathogenesis and pathophysiology of metabolic syndrome can well be understood by studying the molecular mechanisms that control the development and function of adipose tissue. In human body, exist two types of adipose tissue, the white and the brown one, which are reported to play various roles in energy homeostasis. The major and most efficient storage of energy occurs in the form of triglycerides in white adipose tissue while brown adipose tissue actively participates in both basal and inducible energy consumption in the form of thermogenesis. Recent years have observed a rapid and greater interest towards developmental plasticity and therapeutic potential of stromal cells those isolated from adipose tissue. The adipocyte differentiation involves a couple of regulators in the white or brown adipogenesis. Peroxisome proliferators-activated receptor-gamma actively participates in regulating carbohydrate and lipid metabolism, and also acts as main regulator of both white and brown adipogenesis. This review based on our recent research, seeks to highlight the adipocyte differentiation.

Effects of RNA interference targeting HOXA9 on the proliferation and apoptosis of human acute monocytic leukemia U937 cells / 肿瘤研究与临床

Objective To investigate the effects of small interference RNA (siRNA) targeting HOXA9 on the proliferation and apoptosis of human acute monocytic leukemia U937 cell line.Methods Effective and specific siRNA oligo targeting HOXA9 was designed and compounded.It was transfected transiently into U937 cells by cationic liposome.The cells was divided into three groups:experimental group(siRNA targeting HOXA9 was transfected by liposome),negative control group (negative siRNA was transfected by liposome) and cell control group (add equal cells and medium).The expression of HOXA9 mRNA and protein were detected by reverse transcription PCR and Western blot.The cell proliferation was assessed by MTT.The apoptosis of each group were measured by Annexin V-FITC.Results Aftcr transfected by siRNA targeting HOXA9,the relative mRNA expression levels of HOXA9 in the experimental group,negative control group and cell control group were (22.980±0.548) ％,(82.371±1.517) ％ and (84.637±2.252) ％,respectively (P ＜ 0.05),and the relative protein expression levels were (50.377±2.773).％,(105.500±3.900) ％ and (111.392±3.905) ％,respectively (P ＜ 0.05).The inhibitory rates of cell proliferation and the apoptosis rates of the experimental group were significantly increased.The inhibitory rates of cell proliferation of 24 h,48 h and 72 h were (41.909±4.333) ％,(54.470±3.756) ％ and (65.835±1.024) ％,respectively,and the apoatosis rate was (26.800±2.081) ％.Compared with 2 controls,the experimental group differences had statistically significance (P ＜ 0.05).Conclusion siRNA targeting HOXA9 can effectively silence HOXA9 gene expression in U937 cell,suppress cell proliferation and induce cell apoptosis obviously,which providing experimental basis for clinical lenkemia therapy by targeting HOXA9 gene.

CDX gene and tumor / 国际肿瘤学杂志

The caudal-type homeobox gene is a member of homeobox gene—Para-HOX family,including CDX1,CDX2 and CDX4.As a transcriptional factor,CDX plays an important role in embryo development,hematopoietic system formation,intestinal epithelium tissue development and so forth.The abnormal expression of CDX is usually closely related with tumorigenesis.Researching the relationship between CDX and tumors will contribute to tumor diagnosis and treatment.

HOXB5 expression in oral squamous cell carcinoma

Human HOX genes encode transcription factors that act as master regulators of embryonic development. They are important in several processes such as cellular morphogenesis and differentiation. The HOXB5 gene in particular has been reported in some types of neoplasm, but not in oral cancer. OBJECTIVE: The present study investigated the expression of HOXB5 in oral squamous cell carcinoma (SCC) and in non-tumoral adjacent tissues, focusing on verifying its possible role as a broad tumor-associated gene and its association with histopathological and clinical (TNM) characteristics. MATERIAL AND METHODS: RT-PCR was performed to amplify HOXB5 mRNA in 15 OSCCs and adjacent non-tumoral epithelium. A possible association with TNM and histopathologic data was verifed by the chi-square and post-hoc t-test. RESULTS: HOXB5 was amplifed in 60 percent non-tumoral epithelium and in 93.3 percent carcinomas. No statistically signifcant differences were found regarding the HOXB5 mRNA expression and TNM or histological grade. CONCLUSION: HOXB5 is expressed in OSCCs and its role in cancer progression should be further investigated.

Screening and verification of the siRNA targeted for the HOXA10 gene / 肿瘤研究与临床

Objective To obtain effective siRNA fragment of HOXA10 gene and verify its function,to supply experimental evidence for tumor prevention and curation by RNAi targeting to HOXA10 gene.Methods Three pairs of small interfering RNA targeting to the different sites of HOXA10 were designed and introduced into A549.The mRNA expression of HOXA10 of A549 was detected by semi-quantitative RT-PCR,the cell proliferation was assayed by MTT,the apoptosis was measured by flow cytometry.The most effective siRNA assay was screened and was tested the relationship between it and proliferation and apoptosis.Results The mRNA of HOXA10 was inhibited by three siRNAs in A549 cells,among which siRNA1 gave the strongest inhibiting of HOXA10 by ODR was (20.190±1.698) ％.The inhibitory rate of cell proliferation was (69.793±2.092) ％ and the apoptosis rate was (29.593±2.670) ％.Conclusion siRNA1 can specifically degrade HOXA10 mRNA and inhibit the proliferation of A549 cell and promote its apoptosis.

Reversion of multidrug resistance in leukemia K562 cells by RNA interference targeting homeobox A10 gene / 肿瘤

Objective: To investigate a new strategy to reverse multidrug resistance of chronic myeloid leukemia cell line K562 by RNA interference technique through constructing an eukaryotic vector of short hairpin RNA (shRNA) targeting homeobox A10 (HOXA10) gene. Methods: The eukaryotic vector pGPHI/GFP/Neo-HOXA10 with shRNA targeting HOXA10 gene was constructed and then transfected into K562 cells by positive ion liposome. The stable transfectants were vertificated by reverse transcriptase PCR (RT-PCR) 4 weeks after G418 pressure selection. The changes of sensitivity of K562 cells to leurocristine (VCR) and etoposide (VP-16) after transfection with shRNA-HOXA10 were detected by MTT method, and the apoptosis rate was detected by flow cytometry. Results: After selection with G418, the cell clones stably transfected with pGPHI/GFP/Neo-HOXA10 were successfully constructed and verificated by RTPCR. The half inhibitory concentration (IC50) values of VCR and VP-16 for K562 cells transfected with shRNA-HOXA10 were significantly reduced and the apoptosis rate of K562 cells after HOXA10 gene interference combining with chemotherapy was increased significantly as compared with those of shRNA-negative control group and the normal control group (P0.05). Conclusion: The eukaryotic vector of short hairpin RNA (shRNA) targeting HOXA10 gene can enhance the abilities of VCR and VP-16 to inhibit the cell proliferation and induce the apoptosis of K562 cells, namely an increase of sensitivity to these chemotherapeutics in K562 cells. It is hinted that RNA interference targeting HOXA10 gene may reverse the multidrug resistance of leukemia cells in some degree. Copyright© 2011 by TUMOR.

HOXA5 gene and tumor / 国际肿瘤学杂志

As a member of homeobox gene family,HOXA5 is expressed in many organs and can regulate gene expression,cell differentiation and morphogenesis.Its structure and (or) function abnormalities are closely related to the development of leukemia,breast cancer,brain hemangioma,hepatocellular carcinoma and so on.Researching the relationships between HOXA5 and tumors contributes to cancer diagnosis,treatment and prevention.

Expressão do RNA mensageiro de genes HOX em linhagens celulares de carcinomas epidermoides de cabeça e pescoço / HOX genes mRNA expression in head and neck squamous cell carcinoma cell lineages

Os genes HOX são importantes para o controle do desenvolvimento embrionário e regulam aspectos da morfogênese e diferenciação celular. A associação entre os genes HOX e o processo da oncogênese tem sido verificada em casos de carcinomas de cólon, pele, rim, pulmão, mama, leucemias e outros tipos de câncer. O objetivo deste trabalho foi verificar a expressão de genes HOX em linhagens celulares de carcinomas epidermoides de cabeça e pescoço. Para tanto, utilizamos a técnica RT-PCR para analisar a expressão de três grupos de genes HOX em linhagens celulares de carcinomas epidermoides de cabeça e pescoço: HN-6, HN-19, HN-30 e HN-31, utilizando-se os primeiros degenerados HOX1, HOX2 e HOX3 e coamplificação com o gene constitutivo da β-actina. Material de gengiva humana e embrião de camundongo foram utilizados como controle de expressão do gene HOX. Os resultados obtidos mostraram que a expressão dos genes HOX apresentou padrão semelhante aos controles utilizados. Houve, porém, subexpressão do grupo HOX1 nas linhagens HN-6 e HN-19 e superexpressão do grupo HOX-2 na linhagem HN-31. Esses achados nos permitem levantar a hipótese de que os genes HOX podem participar das alterações moleculares observadas em carcinomas epidermoides de cabeça e pescoço.

Baixa estatura por haploinsuficiência do gene SHOX: do diagnóstico ao tratamento / Short stature caused by SHOX gene haploinsufficiency: from diagnosis to treatment

Estudos realizados em pacientes portadores de deleções parciais dos cromossomos sexuais permitiram a caracterização do SHOX, gene localizado na região pseudoautossômica no braço curto dos cromossomos sexuais, fundamental na determinação da altura normal. A perda de uma cópia deste gene na síndrome de Turner (ST) explica dois terços da baixa estatura observada nesta síndrome. A haploinsuficiência do SHOX é detectada em 77 por cento dos pacientes com discondrosteose de Leri-Weill, uma forma comum de displasia esquelética de herança autossômica dominante e em 3 por cento das crianças com baixa estatura idiopática (BEI), tornando os defeitos neste gene a principal causa monogênica de baixa estatura. A medida da altura sentada em relação à altura total (Z da AS/AT para idade e sexo) é uma forma simples de identificar a desproporção corpórea e, associada ao exame cuidadoso do paciente e de outros membros da família, auxilia na seleção de pacientes para o estudo molecular do SHOX. O uso de hormônio de crescimento (GH) está bem estabelecido na ST e em razão da causa comum da baixa estatura com o de crianças com defeitos isolados do SHOX o tratamento destes pacientes com GH é também proposto. Neste artigo será revisado os aspectos clínicos, moleculares e terapêuticos da haploinsuficiência do SHOX.

Studies involving patients with short stature and partial deletion of sex chromosomes identified SHOX gene in the pseudoautosomal region of the X and Y chromosomes. SHOX haploinsufficiency is an important cause of short stature in a diversity of clinical conditions. It explains 2/3 of short stature observed in Turner syndrome (TS) patients. Heterozygous mutations in SHOX are observed in 77 percent of patients with Leri-Weill dyschondrosteosis, a common dominant inherited skeletal dysplasia and in 3 percent of children with idiopathic short stature, indicating that SHOX defects are the most frequent monogenetic cause of short stature. The sitting height/height ratio (SH/H) standard deviation score is a simple way to assess body proportions and together with a careful exam of other family members, effectively selected a group of patients that presented a high frequency of SHOX mutations. Growth hormone treatment of short stature due to TS is well established and considering the common etiology of short stature in patients with isolated defects of SHOX gene, this treatment is also proposed for these patients. Here, we review clinical, molecular and therapeutic aspects of SHOX haploinsufficiency.

Csx/Nkx 2.5 gene expression in embryonic hearts and its mutation in congenital heart disease / 中华围产医学杂志

Objective To explore the Csx/Nkx 2.5 gene expression in the heart during the embryonic period and its mutation in subjects with congenital heart disease(CHD). Methods Immunohistochemistry was used to reveal the Csx/Nkx 2.5 gene expression, and PCR-SSCP-silver staining and DNA sequencing for mutation. Sixty-three embryos or fetus, 126 children with congenital heart diseases and 30 normal controls were included in the study. Results Elevated expression of Csx/Nkx 2.5 gene was found in atrium and trabecular of ventricle. After 16 weeks of gestation, the expression in atrium was stable, while slightly reduced in the trabecular. The expression in the ventricle was lower than that in the atrium in early embryonic stage followed by continuous increase which was most remarkable in 13~16 weeks and kept stable after 16 weeks. No expression of Csx/Nkx 2.5 was detected in epicardium. Three different kinds of gene polymorphisms in the third base of the 21st amino acid codon were found in all subjects:A,G,A/G. Conclusions Gene Csx/Nkx 2.5 plays an important role during the fetal heart development and its expression varies in different parts of the heart during different period in fetal development. Neither the sporadic nor the CHD cases showed any mutations in this study.