Objective： To investigate the proteornic differences between the high-sensitivity （HS） group and low-sensitivity （LS） group of cervical cancer treated by radiotherapy and confirm the radiotherapy sensitivity asso...Objective： To investigate the proteornic differences between the high-sensitivity （HS） group and low-sensitivity （LS） group of cervical cancer treated by radiotherapy and confirm the radiotherapy sensitivity associated proteins in early cervical cancer. Methods： The fresh carcinoma tissues were collected from 10 untreated cervical cancer patients and preserved in the -80 ℃ refrigeratory. The tissues were classified into two groups： high sensitivity group （HS） and low sensitivity group （LS）, according to their response to radiotherapy. In the first part of our experiment, protein separating was performed by using two-dimensional gel electrophoresis （2-DE） with Amersham 18 cm linear pH 3-10 immobilized pH gradient （IPG） strips. The images of the gels were acquired by the scanner and then analyzed by using PD-quest7.3 software to find the differentially expression protein-spots in each group. Then the differentially expressed protein-spots was incised from the gels and digested by trypsin. The peptide mass fingerprintings （PMF） was acquired by matrix assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS） and the proteins were identified by data searching in the Mascot-database. Part of differentially expression proteins were assayed by Western Blot. Results： Most of the gels were clear and successfully analyzed by PD-quest7.3 software. Most of the protein-spots concentrated on the area of 20-100Kda （Mw） and pH4-8. The average number of the protein-spots was 754 ± 64 in HS group and 777 ±48个 in LS group. The match rate was 87.6% between two groups. Five high expression proteins were found in HS group which were low expression in LS group, 3 high expression protein were found in LS group which were low expression in HS group. Reselts of Western Blot were in coincidence to proteomic result. Conclusion： The 2-DE gels image of HS group and LS group with early cervical cancer tissues treated by radiotherapy are successfully acquired. Some differentially expression proteins between the two groups are further confirmed by immunohistochemical assay.展开更多
基金Supported by grants from the Hunan Natural Science foundation (No.06JJ4199)the Hunan Science Technology Foundation (No.2007SK3010)
文摘Objective： To investigate the proteornic differences between the high-sensitivity （HS） group and low-sensitivity （LS） group of cervical cancer treated by radiotherapy and confirm the radiotherapy sensitivity associated proteins in early cervical cancer. Methods： The fresh carcinoma tissues were collected from 10 untreated cervical cancer patients and preserved in the -80 ℃ refrigeratory. The tissues were classified into two groups： high sensitivity group （HS） and low sensitivity group （LS）, according to their response to radiotherapy. In the first part of our experiment, protein separating was performed by using two-dimensional gel electrophoresis （2-DE） with Amersham 18 cm linear pH 3-10 immobilized pH gradient （IPG） strips. The images of the gels were acquired by the scanner and then analyzed by using PD-quest7.3 software to find the differentially expression protein-spots in each group. Then the differentially expressed protein-spots was incised from the gels and digested by trypsin. The peptide mass fingerprintings （PMF） was acquired by matrix assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS） and the proteins were identified by data searching in the Mascot-database. Part of differentially expression proteins were assayed by Western Blot. Results： Most of the gels were clear and successfully analyzed by PD-quest7.3 software. Most of the protein-spots concentrated on the area of 20-100Kda （Mw） and pH4-8. The average number of the protein-spots was 754 ± 64 in HS group and 777 ±48个 in LS group. The match rate was 87.6% between two groups. Five high expression proteins were found in HS group which were low expression in LS group, 3 high expression protein were found in LS group which were low expression in HS group. Reselts of Western Blot were in coincidence to proteomic result. Conclusion： The 2-DE gels image of HS group and LS group with early cervical cancer tissues treated by radiotherapy are successfully acquired. Some differentially expression proteins between the two groups are further confirmed by immunohistochemical assay.
