| Esophageal squamous cell carcinoma is one of the most common malignancies. Up to now, its carcinogenesis mechanism is unclear. It was reported that HPV had related with esophageal carcinoma. However, which role it plays during the malignant transformation of esophageal carcinoma was little known. T-DE, MALDI-TOF-MS, RT-PCR, animal experiment were used to identify and study the differentially expressed proteins between the human immortalized esophageal epithelial cell line (SHEE) transfected by human papillomavirus typelS E6E7 and the malignant transformation cell line (SHEEmt), which was derivated from SHEE. The results could help us to explore the carcinogenesis mechanism of esophageal squamous cell carcinoma that infected with HPV. Materials and methods:1. The proteins were extracted from SHEE and SHEEmt and separated by T-DE, followed coomassie brilliant blue staining. The images of gels were analysed by PDQuest 6.2 software and the differentially expressed proteins were identified by MALDI-TOF-MS.2. The mRNAs of the differentially expressed genes were further analyzed by RT-PCR and the products were cloned and sequenced.3. The differentially expressed proteins were measured between SHEE and SHEEmt cell by western blotting and were detected in esophageal squamous cell carcinoma, adenocarcinoma tissue and normal esophageal tissue by immunohistochemistry.4. The mRNA of differentially expressed genes was blocked by stably expressed antisense RNA to observe the effect on the biological behavior of SHEEmt and. Results:1. Upregulated expressed triosephosphate isomerase (TPI) and fascin 1 were identified in SHEEmt by T-DE and MALDI-TOF- MS.2. The mRNAs of fascin 1 and triosephosphate isomerase were also upregulated in SHEEmt control to SHEE cells, but the mRNA of fascin 1 was more significantly than mRNA of TPI. The amplification sequence of fascin 1 gene was consistent with the sequence of fascin 1 gene in GenBank.3. Upregulated Fascin 1 protein was further assessed in SHEEmt by western blotting. Positive immunoreactions in esophageal squamous cell carcinoma, adenocarcinoma and negative immunoreactions in normal esophageal epithelia cell layer were appeared for fascin 1 by immunohistochemistry. But positive immunoreactions were found in basal cells of epithelia in part of normal tissues.4. pcDNA3.0-fascin expression vector was constructed and transfected into SHEEmt and HeLa cells. SHEEmt -fascincell and HeLa-fascin cell strains were obtained.5. The number of filopodia and microspikes of HeLa-fascin cells was decreased and the cells grew together. The mRNA of fascin 1 was decreased apparently, but fascin 1 protein remained unchanged between transfected cells and untransfected cells.6. The antisense fascin 1 transfectant cells showed no alteration on cell cycle. Fascin I transfectant cells, empty vector transfectant cells and untransfectant cells could develope transplanted tumors in SCID mice.Conclusion:1. TPI and fascin 1 were upregulated expressed in SHEEmt control to SHEE. It indicated that the upregulated expression of fascin 1 and TPI might be involved in the malignant transformation of SHEE to SHEEmt.2. The mRNAs of fascin 1 and TPI were also upregulated in SHEEmt, but the mRNA of triosephosphate isomerase was not upregulated so significantly in SHEEmt.53. Fascin 1 protein was positively stained in tissue of esophageal carcinoma, but it was negative in normal esophageal epithelium by imrnunohistochernical staining. It was further confirmed that fascin 1 should be involved to malignant transformation of esophageal carcinoma.4. Stably expressed antisense fascin 1 RNA has different effects on the morphology of fascin 1 transfectant cells, but didn't affect the division and proliferation of the transfectant cells and untransfectant cells, and transplanted tumors were formed in SCID mice.
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