Molecular targets for liver cancer therapy: From screening of target genes to clinical trials

Cancer arises from the accumulation of genetic alterations, and the inactivation of oncogenes, or recovery of suppressor genes, are promising strategies for cancer treatment. Genome-based drug research starts with identification of target genes and is accomplished by exploitation of target-based drugs such as monoclonal antibodies, small molecules and antisense drugs. Recently, clinical trials for treatment of advanced hepatocellular carcinoma (HCC) have been performed, and the effectiveness of sorafenib, an oral multikinase inhibitor of the vascular endothelial growth factor receptor and Ras kinase, has been demonstrated. In addition to known target genes, microarray technology has enabled us to constitute novel therapeutic targets, and many researchers have applied this technology in studies of HCC and have identified candidate target genes, validated to affect cell growth. In addition, promoter arrays for whole-genome epigenetic aberration analysis, ChIP-chip analysis using tiling arrays, and high-throughput sequencing systems have been applied to drug discovery. To elucidate the status of therapeutic target genes in vivo , development of diagnostic markers for stratification of patients is a pressing need. Here, we review recent advances in microarray technology for liver cancer, discuss the innovations and approaches to therapeutic target discovery, and present data regarding the outcome of gene target therapy using monoclonal antibodies and molecular diagnostic markers in our laboratory.     

风湿性心脏病合并重度肺动脉高压患者体外循环前、后肺血流动力学改变

目的：观察风湿性心脏病（风心病）重度肺动脉高压患者体外循环（ＣＰＢ）前、后肺血流动力学变化规律。方法：体外循环前、后用Ｓｗａｎ－Ｇａｎｚ导管监测１５例风心病重度肺动脉高压患者肺血流动力学参数。结果：ＣＰＢ前肺动脉压为３．７／２．３～１１．６／６．０ｋＰａ（２８／１７～８７／４５ｍｍＨｇ），ＣＰＢ后为２．９／１．１～１１．８／５．５ｋＰａ（２２／８～８９／４１ｍｍＨｇ）。二尖瓣替换术后肺动脉收缩压无明显降低（－９％，Ｐ＞０．０５），肺动脉舒张压（－２０％）和平均压（－１４％）均明显降低（Ｐ＜０．０５）。肺血管阻力（ＰＶＲ）在ＣＰＢ前高达６０．３±４０．９ｋＰａ·ｓ／Ｌ，ＣＰＢ后明显降低（－４０％，Ｐ＜０．０５）。回归方程为：ＣＰＢ后ＰＶＲ＝０．５４６ＰＶＲｂ＋２３．２４，Ｒ２＝０．６５１（ＰＶＲｂ为肺血管阻力基础值）。鱼精蛋白静脉用药１０例，肺动脉压升高者４例；动脉用药５例，均未发现肺动脉压改变。结论：风心病肺动脉高压患者术后大部分ＰＶＲ不能恢复正常。动脉系统用药可减少肝素鱼精蛋白复合物引起的不良反应。     

Chemosensitivity and radiosensitivity profiles of four new human epithelial ovarian cancer cell lines exhibiting genetic alterations in BRCA2, TGFbeta-RII, KRAS2, TP53 and/or CDNK2A

To address the cellular basis for the response to ovarian cancer treatment, we characterized the chemosensitivity and radiosensitivity of four human epithelial ovarian cancer cell lines that harbor different genetic alterations. The TOV-21G, TOV-81D, OV-90, and TOV-112D cell lines were derived from ovarian tumors (TOV) or ascites (OV) from chemotherapy- and radiotherapy-naive patients and were characterized by their mutation spectrum of BRCA2, TGF-RII, KRAS2, TP53, and CDKN2A. Cells were monitored for survival following exposure at various concentrations to different cytotoxic agents including cisplatin, camptothecin or paclitaxel or to different doses of -irradiation. At the lowest doses, the TGF-RII-mutated and KRAS2-mutated cell line, TOV-21G, and the BRCA2-mutated cell line, TOV-81D, demonstrated a significantly higher sensitivity to cisplatin and -irradiation than the TP53-mutated cell lines, TOV-112D and OV-90. At higher doses, differences between the TP53-mutated lines were observed with TOV-112D being less sensitive to cisplatin than OV-90 that also harbors a CDNK2A mutation. All cell lines were similarly sensitive to high doses of -irradiation. In contrast, sensitivity to camptothecin or paclitaxel was not significantly different between all cell lines, irrespective of the mutation status of BRCA1, BRCA2, TGF-RII, KRAS2, TP53, and CDKN2A. The observed responses to treatment are consistent with the current knowledge concerning BRCA2, TGF-RII, KRAS2, TP53, and/or CDKN2A aberrant function.The first and second authors contributed equally to this work.     

肺癌组织抑癌基因P53异常分析

Ｐ５３基因５－８外显子ＰＣＲ产物的ＤＮＡ序列分析及石蜡包埋组织Ｐ５３鼠抗人单克隆抗体（Ｄｏ－１）免疫组化分析发现：５７例原发性肺癌中３７例有Ｐ５３蛋白积累。ＤＮＡ／ＰＣＲ产物直接测序发现：９／１２例小细胞肺癌（ＳＣＬＣ），８／１５非小细胞肺癌（ＮＳＣＬＣ）Ｐ５３基因突变，错义突变为１２／１７，Ｇ→Ｔ突变为６／１７。免疫组化发现：１１／１９例ＳＣＬＣ，１９／３７例ＮＳＣＬＣ Ｐ５３蛋白染色阳性。４／     

Scanning Microscopy of Colonic Mucin during Carcinogenesis: Is It Clinically Applicable?

The integrity of the colonic mucin layer has been reported to be altered during carcinogenesis in both humans and rodents. Prior to attempting scanning microscopic techniques on colonic mucosa of patients at high risk to develop colorectal cancer, these procedures were performed on colonic mucosa from rats with chemically induced colon cancers. Substantial technical difficulties in preparation and serious subjectivity in interpretation of the scanning micrographs were encountered. The major technical problem was the unpredictable retention of the mucin layer upon both normal and cancerous mucosae. Visual interpretation of the integrity or disruption of the mucin layer with the scanning electron microscope revealed variable fenestration and fraying of the mucin in both normal and cancerous colons. Our findings suggest that scanning electron microscopy of colonic mucin may not be a reliable screening procedure for (pre)cancerous changes in human colonic mucosae.