Alternative splicing of pre-mRNA is an important mechanism for regulating gene function at the post-transcription level and for producing proteomic diversity in higher eukaryotes. The alternative splicing is regulated by the interaction between diverse cis-acting elements and trans-acting factors. Alternative splicing events of oncogenes, tumor suppressor genes and metastasis suppressor genes are associated with the initiation and development of human neoplasms. The protein isoforms sourced from alternative splicing take part in regulating the gene transcription, cell cycle, apoptosis of cells, and playing a role in tumor growth. It is possible for molecular therapy to target directly isoforms of protein produced by alternative splicing or to interfere with the process of alternative splicing.
Alternative Splicing ; genetics ; Humans ; Neoplasms ; genetics ; RNA Precursors ; metabolism ; RNA, Neoplasm ; analysis ; Transcription, Genetic

Actinomyces meyeri is rarely isolated in cases of actinomycosis. The identification of A. meyeri had historically been difficult and unreliable. With the recent development of 16S ribosomal RNA (16S rRNA) sequencing, Actinomyces species such as A. meyeri can be isolated much more reliably. A. meyeri often causes disseminated disease, which can be secondary to frequent pulmonary infections. A penicillin-based regimen is the mainstay of A. meyeri treatment, with a prolonged course usually required. Here, we report a case of pulmonary actinomycosis with brain abscess caused by A. meyeri that was initially thought to represent lung cancer with brain metastasis.
Actinomyces* ; Actinomycosis ; Brain Abscess ; Brain* ; Lung Neoplasms ; Lung* ; Neoplasm Metastasis ; RNA ; RNA, Ribosomal, 16S ; Sequence Analysis

OBJECTIVEEstablishment of a gene expression profile associated with differentiation inducing the glioma cells was made possible.

METHODThe expression level of 18 000 genes in glioma cells was evaluated before and after induction with sodium phenyl-butyrate for 2 hours or 6 days by cDNA array technique, with the results proved by multi-dot blot.

RESULTSNinety-eight gene expressions in the glioma cells were changed after the induction, with some genes in transcription and translation systems down-regulated, some oncogenes down-regulated, and some differentiation or apoptosis genes up-regulated. Eighteen unknown EST fragments were changed also.

CONCLUSIONA gene expression profile associated with differentiation-inducing the glioma cells including 98 genes has been established.

Cell Differentiation ; DNA, Neoplasm ; analysis ; Gene Expression ; Gene Expression Profiling ; Glioma ; genetics ; metabolism ; pathology ; Humans ; Oligonucleotide Array Sequence Analysis ; RNA, Neoplasm ; analysis

Synovial sarcoma (SS) is a rare high grade soft tissue sarcoma with unusual epithelial differentiation. SS does not arise from synovial tissue, but rather originates from pluripotential mesenchymal cells. Three histomorphologic subtypes of SS have been described: biphasic, monophasic, and poorly differentiated. Typically, biphasic SS is composed of epitheloid cells forming pseudoglandular spaces and spindle cells resembling fibrosarcoma. We report a rare case of a 65-year-old man who presented with two asymptomatic masses on his abdomen and axillary regions. The pertinent histopathologic, immunohistochemical, cytogenetic analysis and radiographic examinations represented monophasic fibrous SS with multiple visceral metastasis. We also found SYT-SSX fusion gene transcript specific for SS using RNA samples taken from the lesional tissues.
Abdomen ; Aged ; Cytogenetic Analysis ; Fibrosarcoma ; Humans ; Neoplasm Metastasis* ; RNA ; Sarcoma ; Sarcoma, Synovial*

BACKGROUNDTo investigate the differential expression levels of thymosin beta 10 (T beta 10) and the corresponding changes of actin filament organization in human tumor cell lines with different metastatic potential.

METHODSFour groups of nine human tumor cell lines with different metastatic potential were analyzed for the amount of T beta 10 mRNAs by Northern blot and for their peptide expression levels by immunohistochemistry. The filamentous actin (F-actin) was observed by staining of TRITC-phalloidin to detect changes in actin organization.

RESULTSIn comparison with non-/weakly metastatic counterparts, T beta 10 was upregulated in highly metastatic human lung cancer, malignant melanoma and breast cancer cell lines. Staining of TRITC-phalloidin revealed less actin bundles, a fuzzy network of shorter filaments and some F-actin aggregates in the highly metastatic tumor cells. Meanwhile, the actin filaments were robust and orderly arranged in the non-/weakly metastatic cancer cell lines.

CONCLUSIONT beta 10 levels correlate positively with the metastatic capacity in human tumors currently examined. The increasing metastatic potential of tumor cells is accompanied by a loss of F-actin, poorly arranged actin skeleton organizations and presence of F-actin aggregates. There is a consistent correlation between the elevated T beta 10 expression and the disrupted actin skeleton.

Actin Cytoskeleton ; ultrastructure ; Blotting, Northern ; Cell Line, Tumor ; Humans ; Immunohistochemistry ; Neoplasm Metastasis ; genetics ; ultrastructure ; RNA, Messenger ; analysis ; Thymosin ; analysis

BACKGROUNDIn humans telomerase is expressed in most cancers and immortal cell lines, and activation of telomerase may play important roles in tumorigenesis and immortalization. This study was to investigate the roles of telomerase activity (TA) and human telomerase RNA (hTR) in sebaceous carcinoma of the eyelid.

METHODSThe telomerase repeated amplification protocol (TRAP) was used to demonstrate telomerase activity in 12 cases of sebaceous carcinoma of the eyelid. In situ hybridization (ISH) was used to demonstrate the expression of hTR in 55 cases of paraffin-embedded sebaceous carcinoma of the eyelid, and the results were compared with the proliferative index determined by Mib-1 immuno-labeling, histological patterns and recurrence of the tumor.

RESULTSDifferent telomerase activity was shown in the 12 cases of sebaceous carcinoma of the eyelid. The positive expression of hTR was 85.5% (47/55) in tumor cells, but not in the adjacent tissues. The positive expression of hTR was correlated with the proliferative activity (as assessed by Mib-1 immunolabelling, r = 0.942, P < 0.001) and the differentiation of sebaceous carcinoma of the eyelid (chi(2) = 17.621, P < 0.001), but not significantly related to tumor recurrence. The level of hTR expression increased with the decrease of differentiation of sebaceous carcinoma of the eyelid.

CONCLUSIONSThe results suggest that the up-regulation of telomerase expression plays some roles in tarsal gland carcinogenesis, and the expression of hTR is a useful marker for malignant degree of sebaceous carcinoma of the eyelid.

Biomarkers, Tumor ; analysis ; Eyelid Neoplasms ; enzymology ; pathology ; Humans ; In Situ Hybridization ; Neoplasm Recurrence, Local ; enzymology ; RNA ; analysis ; Sebaceous Gland Neoplasms ; enzymology ; pathology ; Telomerase ; analysis

Acid Anhydride Hydrolases ; Carcinoma, Hepatocellular ; etiology ; genetics ; Humans ; Liver Neoplasms ; etiology ; genetics ; Mutation ; Neoplasm Proteins ; genetics ; RNA, Messenger ; analysis

OBJECTIVESTo observe the expression of betaig-h3 in normal cornea and keratoconus and to elucidate the role of extracellular matrix in keratoconus.

METHODSIn situ hybridization was used to detect the expression of betaig-h3 in the cornea. The cDNA library was screened with human betaig-h3 cDNA probe to locate betaig-h3 mRNA in cells.

RESULTSExpression of betaig-h3 was found mainly in the stroma of the normal cornea and keratoconus, but decrease depending on the degree of keratopathy. In some serious cases, no expression signal was detected. The strongest expression was seen at the border of the normal region and keratoconus.

CONCLUSIONSbetaig-h3, the structural component of the extracellular matrix, can affect cell adhensiveness in the development of corneal fibrous interstitial organization. During the development of keratoconus, decreasing levels of betaig-h3 cause the diminution of corneal steadiness, which is related to formation of keratoconus.

Cornea ; metabolism ; Extracellular Matrix Proteins ; Humans ; Keratoconus ; metabolism ; Neoplasm Proteins ; genetics ; RNA, Messenger ; analysis ; Transforming Growth Factor beta ; Wound Healing

BACKGROUND: Differentiated thyroid cancer is the most common endocrine malignancy. Despite advances in the treatment of thyroid cancer, disease recurrence and metastasis may occur in as many as 20% of patients, and so continues to pose major problems in its clinical management. Serum thyroglobulin (Tg) measurements, by immunoassay, are used to detect residual or recurrent thyroid cancer following thyroid ablation. However, the usefulness of immunoassay is limited by both the requirement for thyroid hormone withdrawal, to attain optimal test sensitivity, and interference by the antithyroglobulin antibody (Anti-Tg Ab). Recent studies have reported the clinical usefulness of reverse transcription-polymerase chain reaction (RT-PCR) detection of Tg mRNA in the peripheral blood of patients with differentiated thyroid carcinomas. We performed this study to evaluate the usefulness RT-PCR of Tg mRNA in peripheral blood of patients with thyroid carcinoma following a total thyroidectomy and radioiodine ablation therapy. METHODS: Forty cases that underwent a total thyroidectomy and radioiodine ablation therapy were included in this study. Of the 40 patients, 35 were papillary carcinomas and 5 were follicular carcinomas. Ten normal control subjects were also studied. Tg mRNA was extracted. Then RT-PCR, and nested RT-PCR, were run with specific Tg primers. Concurrently, DNA sequencing of the isolates was carried out to prove the isolates were identical to the nucleotide sequence of the Tg. RESULTS: The Tg was detected in 4 of 19 patients, with either a residual thyroid bed, or metastasis, on a 131I whole body scan and in 1 of 21 patients with a negative radioiodine scan. Surprisingly, the Tg mRNA was detected in all the patients and normal controls. CONCLUSION: From our results we can not recommend Tg mRNA, detected by RT-PCR in peripheral blood, as a tumor marker superior to that of the Tg serum level. We consider an intensive re-evaluation of the method is required before considering its clinical applications.
Base Sequence ; Carcinoma, Papillary ; Diagnosis* ; Humans ; Immunoassay ; Neoplasm Metastasis ; Recurrence ; RNA* ; RNA, Messenger ; Sequence Analysis, DNA ; Thyroglobulin* ; Thyroid Gland* ; Thyroid Neoplasms* ; Thyroidectomy ; Whole Body Imaging

OBJECTIVETo investigate the effect of small interference RNA (siRNA) on mdr1 and P-glyco-protein (P-gp) expression of multi-drug resistance (MDR) human leukemia cell line K562/A02.

METHODSThree si RNAs (si-mdr1-1, si-mdr1-2, si-mdr1-3) which were specifically targeted mdr1 gene were synthesized and transfected into K562/A02 cells. Expression of mdr1 mRNA was assayed by RT-PCR. P-gp expression and intracellular daunorubicin (DNR) concentration were determined by flow cytometry. 50% inhibition concentration (IC(50)) of doxorubicin (ADM) on K562/A02 was determined by MTT method.

RESULTSTreatment of K562/A02 cell with the 3 kinds of siRNAs resulted in a reversal of MDR of a different extent. The third siRNA was more effective in the suppression of mdr1 with a significant reduction of (58.0 +/- 1.54)% of the mdr1 mRNA expression. Positive expression rate of p170 decreased from (76.0 +/- 1.0)% to (19.6 +/- 1.9)%, and the relative efficiency of K562/A02 to ADM was 70.4%. The intracellular accumulation of DNR increased after siRNA treatment.

CONCLUSIONThe siRNA could effectively restore the sensitivity of K562/A02 cells to conventional chemotherapeutic agents.

Base Sequence ; Daunorubicin ; pharmacokinetics ; Drug Resistance, Neoplasm ; Genes, MDR ; physiology ; Humans ; K562 Cells ; drug effects ; Molecular Sequence Data ; RNA, Messenger ; analysis ; RNA, Small Interfering ; pharmacology