卵巢癌相关基因扩增的实验研究

目的我们用卵巢癌SKOV_3细胞株,对裸鼠进行转瘤、应用核酸杂交技术,对肿瘤组织erbB-2、myc、ras基因的扩增进行检测,分析其扩增对卵巢癌的治疗、预后的影响,为卵巢癌的治疗、预后提供实验依据。方法将24只探鼠注射SKOV_3（1×107／ml）,传瘤16天。将传它探鼠分为对照组,干扰素治疗组、维甲酸治疗组、干扰素十维甲酸联合治疗组,每组6只裸鼠。治疗4周后,取肿瘤组织.按常规方法提取肿瘤组织DNA,用~35S它标记的c-myc、ras、erbB-2探针进行Southern杂交,放射自显影3天洗片。结果经用光密度扫描仪扫描,对照组基因的DNA含量最高,干扰素组和维甲酸组校对照组低,联合应用组基因DNA含量最低。结论本实验应用SKOV_3细胞,对裸鼠转瘤后,应用干扰素及维甲酸联合诱导治疗,c-myc、ras、erbB-2癌基因的扩增明显下降,肿瘤好转,为临床卵巢癌的诱导分化治疗提供了实验依据、但对其抗肿瘤的确切机理仍需要进一步研究,也需临床治疗的进一步验证。     

卵巢癌诱导分化治疗后相关基因表达的实验研究

许多研究表明，癌基因扩增与表达异常在恶性肿瘤的发生和演变中发挥重要作用。与肿瘤的预后有密切的关系，我们用卵巢癌SKOV3细胞株，对探鼠进行转瘤、应用核酸杂交技术，对肿瘤组织的erbB-2、c-myc、H-ras基因的mRNA进行检测和分析，结果显示单独应用干扰素、维甲酸治疗，三种癌基因的表达下降；联合用药基因表达的下降更明显。表明癌基因的表达与肿瘤的治疗、预后密切相关。     

脂质体-c-erbB-2反义寡核苷酸对人卵巢癌裸鼠皮下移植瘤的治疗作用

目的　探讨c erbB 2反义寡脱氧核苷酸 (ASODN )对人卵巢癌裸鼠皮下移植瘤的治疗作用。方法　将 15只裸鼠建立人卵巢癌皮下移植瘤模型 ,然后随机分成 3组予以不同条件处理 :对照组 (腹腔注射转染液和脂质体 ) ,正义治疗组 (腹腔注射脂质体 c erbB 2 -SODN ) ,反义治疗组 (腹腔注射脂质体 c erbB 2 -ASODN )。治疗期间定期观测裸鼠体重和肿瘤体积 ,计算抑瘤率及肿瘤缩小率。利用RT -PCR技术比较治疗后各组肿瘤组织中c erbB 2基因水平。结果　反义治疗组与正义治疗组的抑瘤率分别为 71.2 %和 7.6% ,反义治疗组肿瘤缩小率为 2 2 .7% ,与正义治疗组相比较有显著性差异。各组裸鼠体重变化无明显差异。RT -PCR结果显示反义治疗组c erbB 2表达水平降低约 61.0 %。结论　脂质体 c erbB 2 -ASODN对人卵巢癌裸鼠皮下移植瘤有一定的治疗作用 ,可能成为日后卵巢癌基因治疗的重要途径。     

siRNA介导的间皮素基因沉默对卵巢癌裸鼠腹腔移植瘤生长的影响

目的:研究间皮素(mesothelin, MSLN)基因被沉默后的卵巢癌细胞SKOV3-MSLN-shRNA增殖能力的变化,以及MSLN基因沉默对裸鼠卵巢癌腹腔移植瘤生长的影响.方法:利用针对MSLN基因的siRNA慢病毒液和对照组慢病毒液分别感染SKOV3细胞,建立SKOV3-MSLN-shRNA细胞和SKOV3-MSLN-neg细胞的稳定转染株;以SKOV3细胞作空白对照,用平板克隆形成法和细胞计数法检测3组细胞的增殖能力.应用3组细胞分别建立裸鼠卵巢癌腹腔移植瘤模型,2周后处死裸鼠,观察每组动物的成瘤率及每只裸鼠的肿瘤质量、肿瘤数目和肿瘤种植部位等.结果:SKOV3-MSLN-shRNA细胞的体外增殖能力明显低于SKOV3-MSLN-neg细胞和SKOV3细胞,差异有统计学意义(P<0.05).SKOV3-MSLN-shRNA组裸鼠成瘤率为60%,而2个对照组SKOV3-MSLN-neg和SKOV3细胞的裸鼠成瘤率均为100%,差异无统计学意义(P＞0.05).SKOV3-MSLN-shRNA组动物的肿瘤数目、肿瘤质量及肿瘤种植部位明显低于2个对照组 (P<0.05).结论:MSLN基因沉默可以降低卵巢癌SKOV3细胞的体外增殖能力,同时可以抑制卵巢癌裸鼠腹腔移植瘤的生长和种植.     

α-干扰素和维甲酸对人卵巢癌细胞的联合抗增殖作用

采用台盼益拒染法及3H-脱氧胸苷和3H-亮氨酸掺入实验，观察α-干扰素和维甲酸联合对人卵巢癌SKOV3细胞DNA及蛋白质合成的影响。结果显示：1μm／L维甲酸和1000μ／mlα-干扰素联合可显著降低SKOV3细胞的增殖速度，抑制其DNA和蛋白质的合成，联合作用强于二者单独应用之和，且细胞毒性并不增加。提示小剂量维甲酸和α-干扰素联合可用于卵巢癌的治疗。     

四硫化四砷对裸鼠卵巢癌皮下移植瘤生长的影响

目的 利用BALB／C裸鼠和人卵巢癌细胞株SKOV3建立裸鼠皮下移植瘤,探讨四硫化四砷在卵巢癌组织中的抑瘤作用及机制。方法 建立卵巢癌SKOV3裸鼠皮下移植瘤模型,按照给药剂量（50mg／kg和100mg／kg）及肿瘤生成大小（3mm×3mm和10mm×10mm）将裸鼠随机分成小剂量早期治疗组、大剂量早期治疗组、小剂量治疗组和大剂量治疗组,分别予四硫化四砷隔日灌胃,并设裸鼠生理盐水对照组。观察四硫化四砷对肿瘤的生长的影响;RT-PCR法及Western blotting法检测四硫化四砷对肿瘤组织中Bcl-2／Bax蛋白和mRNA表达的影响。结果 各组裸鼠一般情况良好。小剂量早期治疗组、大剂量早期治疗组和大剂量治疗组的抑瘤率分别为84.98％、87. 55％和74.25％,与小剂量治疗组的35.19%相比,差异均有统计学意义（P＜0.05）。RT-PCR显示肿瘤组织Bcl-2 mRNA表达下调和Bax mRNA表达上调,Western blotting显示其Bcl-2蛋白表达降低和Bax蛋白表达增加。结论 四硫化四砷可以抑制卵巢癌裸鼠皮下移植瘤的生长,并诱导卵巢癌细胞凋亡。其作用机制可能与凋亡调控基因Bcl-2／Bax表达的改变有关。     

妇科（080435～080462）

反义HIF-1α基因联合血管抑素基因对人卵巢癌裸鼠移植瘤的抑制作用；CIK细胞联合顺铂对卵巢癌耐药细胞SKOV3/CDDP的杀伤作用；葡萄糖转运体p63和DNA蛋白激酶在卵巢浆液性肿瘤中的表达及其意义；mIL-12基因转染人卵巢癌SKOV3细胞株抗肿瘤免疫作用研究；二维及彩色多普勒超声鉴别卵巢肿瘤228例。     

妇科

D卵巢原发癌与转移癌中survivin mRNA和caspase-3 mRNA的临床研究,乙酰肝素酶基因转染SKOV3细胞对细胞增殖、粘附、侵袭能力的影响,Kilon基因在卵巢上皮性肿瘤中的表达状况的探讨,抗HER-2工程抗体chA21和Herceptin对人卵巢癌SKOV3裸小鼠移值瘤的抑制作用,ERK信号转导通路蛋白在上皮性卵巢肿瘤组织中的表达及与肿瘤恶性程度的相关性研究,内皮抑素基因联合重组干扰素-α蛋白对卵巢癌抑制作用的研究,卵巢癌6811抗独特型微抗体诱导抗肿瘤免疫应答的体外实验,端粒酶抑制剂对顺铂不同敏感性卵巢癌细胞株的影响,     

间皮素siRNA慢病毒对卵巢癌裸鼠移植瘤的治疗效果

目的： 探讨间皮素（mesothelin,MSLN）siRNA慢病毒对人卵巢癌裸鼠腹腔移植瘤的治疗作用。 方法： 利用人卵巢癌SKOV3细胞建立卵巢癌裸鼠腹腔移植瘤模型。实验分为3组：治疗组、阴性对照组和空白对照组。治疗组应用携间皮素siRNA慢病毒液(LV-MSLN-shRNA)对裸鼠模型进行治疗,阴性对照组采用空载体(LV-MSLN-neg)慢病毒液进行治疗,空白对照组注射PBS治疗。从裸鼠生长一般情况、成瘤率、体质量差、瘤体质量和肿瘤转移部位数等方面观察治疗效果, Western blotting法检测移植瘤组织中间皮素蛋白的表达情况。 结果： 用间皮素siRNA慢病毒液对荷瘤裸鼠进行注射的治疗组裸鼠的成瘤率、体质量差、瘤体质量及肿瘤形成部位数均明显低于空白对照组和空载体LV-MSLN-neg治疗组（P<0.01）。注射慢病毒液后瘤体组织中间皮素蛋白表达明显降低（P<0.05）。 结论： 间皮素siRNA慢病毒液对卵巢癌腹腔移植瘤的生长有显著抑制作用。     

MiRNA-210通过PI3/AKT信号通路上调E2F3促进卵巢癌SKOV3细胞增殖作用

摘 要：［目的］ 研究miRNA-210是否通过PI3/AKT信号通路上调E2F3促进卵巢癌SKOV3细胞的增殖作用。［方法］ 收集24例卵巢癌患者的临床组织。QRT-PCR分别检测肿瘤组织和癌旁组织中miRNA-210的表达;Western blot检测肿瘤组织和癌旁组织中AKT、P-AKT、E2F3表达;CCK-8检测三组细胞的增殖能力;平板克隆形成实验检测三组细胞的克隆形成能力;裸鼠皮下荷瘤模型检测三组细胞的成瘤能力。［结果］ 肿瘤组织中miRNA-210的表达高于癌旁组织;肿瘤组织中P-AKT 和E2F3的表达高于癌旁组织;转染结束后,三组细胞中过表达载体miRNA-210转染SKOV3细胞中miRNA-210表达明显高于对照组和空白组,而对照组和空白组之间无差异;CCK-8检测结果显示miRNA-210过表达后促进了SKOV3细胞增殖;平板克隆形成实验结果显示miRNA-210过表达后SKOV3细胞克隆形成能力明显增强;裸鼠皮下荷瘤模型结果显示miRNA-210过表达后SKOV3细胞皮下成瘤能力明显强于对照组和空白组;Western blot结果显示miRNA-210过表达后P-AKT和E2F3表达增加,但是miRNA-210过表达SKOV3细胞系中加入1μmol/L Wortmannin后E2F3表达随之下调。［结论］ MiRNA-210通过PI3/AKT信号通路上调E2F3,从而促进卵巢癌SKOV3细胞增殖作用。     

Occupational and environmental radiation and cancer

Epidemiologic evidence on the relation between occupational and environmental radiation and cancer is reviewed. Studies of pioneering radiation workers, underground miners, and radium dial painters revealed excess cancer deaths and contributed to the setting of radiation protection standards and to theories of carcinogenesis. Occupational exposures today are generally much lower than in the past, thus any associated increases in cancer will be difficult to detect. Pooling investigations of these more recently exposed workers, however, has the potential to validate current estimates of risk used in radiation protection. New information on the effects of chronic radiation exposure also may come from studies in the former Soviet Union of Chernobyl clean-up workers and of workers at the Mayak nuclear facilities. Studies of environmental radiation exposures, other than radon, are largely inconclusive, due mainly to the difficulties in detecting the low risks associated with low dose exposures. Thyroid cancer, however, has been linked to environmental radiation from the Chernobyl accident and from nuclear weapons tests. Low-level radiation released during normal operations at nuclear plants has not been found to increase cancer rates in surrounding populations. Radon, a human carcinogen, is the most ubiquitous exposure to human populations; remediating high residential-radon levels is recommended, recognizing that the exposure can never be removed completely because it occurs naturally.     

New and emerging radiosensitizers and radioprotectors

The combination of chemotherapy and radiation has led to clinical breakthroughs in several disease sites, and current work continues to define optimum combinations of proven chemotherapy as well as more recently available, noncytotoxic agents. Administration of systemic therapies allows modulation of radiation response to improve tumor control (radiosensitization) or to prevent normal tissue toxicity (radioprotection). Substantial progress has been made in identifying the targets of standard chemotherapeutic radiation sensitizers and protectors as well as in the introduction of a new generation of molecularly targeted therapies in combination with radiation. We have reviewed the most recent, predominantly early phase clinical trials combining systemic agents with radiation. Although the proof of an improved schedule ultimately needs to come from well-run Phase III trials, the search among schedules could be shortened by the use of surrogate endpoints such as presence of active drug metabolites in the tumor. This has been accomplished only in a few cases and needs to become a more standard part of radiation sensitizer and protector trials.     

Vitamin D and melanoma and non-melanoma skin cancer risk and prognosis: A comprehensive review and meta-analysis

Vitamin D is formed mainly in the skin upon exposure to sunlight and can as well be taken orally with food or through supplements. While sun exposure is a known risk factor for skin cancer development, vitamin D exerts anti-proliferative and pro-apoptotic effects on melanocytes and keratinocytes in vitro. To clarify the role of vitamin D in skin carcinogenesis, we performed a review of the literature and meta-analysis to evaluate the association of vitamin D serum levels and dietary intake with cutaneous melanoma (CM) and non-melanoma skin cancer (NMSC) risk and melanoma prognostic factors. Twenty papers were included for an overall 1420 CM and 2317 NMSC. The summary relative risks (SRRs) from random effects models for the association of highest versus lowest vitamin D serum levels was 1.46 (95% confidence interval (CI) 0.60–3.53) and 1.64 (95% CI 1.02–2.65) for CM and NMSC, respectively. The SRR for the highest versus lowest quintile of vitamin D intake was 0.86 (95% CI 0.63–1.13) for CM and 1.03 (95% CI 0.95–1.13) for NMSC. Data were suggestive of an inverse association between vitamin D blood levels and CM thickness at diagnosis. Further research is needed to investigate the effect of vitamin D on skin cancer risk in populations with different exposure to sunlight and dietary habits, and to evaluate whether vitamin D supplementation is effective in improving CM survival.     

Religiosity and Physical and Emotional Functioning Among African American and White Colorectal and Lung Cancer Patients

The literature suggests that religiosity helps cope with illness. The present study examined the role of religiosity in functioning among African Americans and Whites with a cancer diagnosis. Patients were recruited from an existing study and mailed a religiosity survey. Participants (N = 269; 36% African American, 56% women) completed the mail survey, and interview data from the larger cohort was utilized in the analysis. Multivariate analyses indicated that in the overall sample religious behaviors were marginally and positively associated with mental health and negatively with depressive symptoms. Among women, religious behaviors were positively associated with mental health and negatively with depressive symptoms. Religiosity was not a predictor of study outcomes for men. Among African Americans, religious behaviors were positively associated with mental health and vitality. Among Whites, religious behaviors were negatively associated with depressive symptoms. These findings suggest a mixed role of religious involvement in cancer outcomes. The current findings may have applied potential in the areas of emotional functioning and depression.     

Fruit and vegetables and cancer risk

The possibility that fruit and vegetables may help to reduce the risk of cancer has been studied for over 30 years, but no protective effects have been firmly established. For cancers of the upper gastrointestinal tract, epidemiological studies have generally observed that people with a relatively high intake of fruit and vegetables have a moderately reduced risk, but these observations must be interpreted cautiously because of potential confounding by smoking and alcohol. For lung cancer, recent large prospective analyses with detailed adjustment for smoking have not shown a convincing association between fruit and vegetable intake and reduced risk. For other common cancers, including colorectal, breast and prostate cancer, epidemiological studies suggest little or no association between total fruit and vegetable consumption and risk. It is still possible that there are benefits to be identified: there could be benefits in populations with low average intakes of fruit and vegetables, such that those eating moderate amounts have a lower cancer risk than those eating very low amounts, and there could also be effects of particular nutrients in certain fruits and vegetables, as fruit and vegetables have very varied composition. Nutritional principles indicate that healthy diets should include at least moderate amounts of fruit and vegetables, but the available data suggest that general increases in fruit and vegetable intake would not have much effect on cancer rates, at least in well-nourished populations. Current advice in relation to diet and cancer should include the recommendation to consume adequate amounts of fruit and vegetables, but should put most emphasis on the well-established adverse effects of obesity and high alcohol intakes.     

肾上腺素能β受体与肿瘤生长及转移

大量研究表明肿瘤细胞可表达β受体，而一些神经递质、药物和社会心理因素可能通过β受体影响肿瘤的生长和转移，β受体激动剂、β受体阻滞剂以及抑郁等社会心理因素可加强或削弱这种作用。这为表达β受体肿瘤的治疗开辟了新的道路，提供了新的治疗靶点。     

VEGF及KDR在大肠腺瘤和腺癌中的表达及意义

目的：探讨VEGF和KDR在大肠腺瘤和大肠腺癌中的表达及临床病理特征的关系。方法：大肠腺瘤和大肠腺癌组织标本各100例,采用免疫组织化学染色法检测VEGF和KDR在标本中的表达情况。结果：VEGF和KDR在大肠腺癌组中的阳性表达明显高于大肠腺瘤组（P〈0.05）;在正常大肠黏膜均未见VEGF和KDR表达的阳性染色;VEGF阳性表达组中KDR的阳性表达率为70%,显著高于VEGF阴性表达组中KDR的阳性表达率16%,两组比较有统计学意义（P〈0.01）。结论：大肠腺癌组织中KDR的表达与肿瘤大小、转移情况、浸润深度密切相关;VEGF和KDR在大肠腺瘤中的表达与患者的年龄、性别及分型均无相关性,而与增生程度相关（P〈0.05）。在大肠腺癌患者中VEGF及KDR表达更高,二者具有协同效应。     

Cell and tissue interactions in carcinogenesis and metastasis and their clinical significance

This review describes a new vision for future directions in the study of metastatic cancer biology and pathology. It is based upon clinical and experimental observations on the constituent cell lineages within a neoplasm and on tumour-host interactions. The vision incorporates information from studies in population biology, developmental biology and experimental pathology as well as investigations upon human malignant disease. The assembled information reveals that invasion and metastasis are supra-cellular manifestations of "emergent behavior" among combinations of normal and malignant cell lineages in vivo. Emergent behavior is a combinatorial interactive process in which a population displays new traits which cannot be achieved by individuals acting separately and which subside when the specific population mix disaggregates. Disruption of such pathological interactions in the field of a developing primary or secondary tumour is, therefore, required to disable the malignant population and arrest progression without tissue destruction. These conclusions originate, in part, from principles which govern the sociobiology and group behavior of bees, ants, fish, birds and human societies. In all these social organisms, external factors can disrupt signaling mechanisms and induce expanding self-perpetuating rogue behavior, leading to social disintegration. These principles also apply to cellular societies composing higher animals, which likewise need intrinsic rules to maintain social order and avoid anarchy, and recognition of this is essential for advancing future research on the mechanisms involved in carcinogenesis and metastasis. Summarised evidence is presented here to support the conclusion that miscommunications between cells and tissues in the region of the developing tumour and its metastases are the main direct perpetrators of malignant disease. Genetic lesions (mutations, deletions, translocations, reduplications, etc.), commonly seen in cancers, can significantly disrupt important molecular pathways in the networks of communications needed to sustain orderly tissue/organ structure and function. However, genetic lesions can also, themselves, be induced by abnormal cell interactions initiated by extrinsic carcinogenic agents such as chemicals, viruses, hormones and radiation. The evidence shows that, irrespective of the initiating cause, it is this miscommunication in the region of a developing tumour and its metastases that is ultimately responsible for the emergence and progression of the disease. The article describes how this information collectively, provides a framework for designing specific novel therapeutic approaches targeting the cell and tissue interactions driving tumour metastasis and its manifold effects on the whole body.