| Objective: The current paper aims to study and analyze the specific tumor markers in the serum of patients with ovarian cancer,to identify the relevant protein in ovarian cancer as well as to confirm the specific markers in ovarian tumors,hopefully,all of which will be able to offer guidance to clinical diagnosis.Method: The study selected 15 patients who were diagnosed with ovarian cancer from August,2016 to February,2017.Their pathologic and clinical data are complete.Another 15 healthy women were chosen as the reference group.Four milliliter of venous blood was drawn from all the subjects to conduct the following experiments: two-dimensional gel electrophoresis,liquid chromatography-mass spectrometry(LC-MS),enzyme linked immunosorbent assay(ELISA)and protein immunoblotting,through which the differences of the relevant protein in the serum between healthy women and patients with ovarian cancer were compared and contrasted.Results:(1)By contrasting the experimenting results of the two groups of women in the two-dimensional gel electrophoresis,the study finds that among all the subjects with ovarian cancer,the content of serum protein in three to four groups is significantly more than that of the healthy women.The most obvious node shows abnormal increase inprotein spot,with its molecular weight of the protein reaching 45 KD and its isoelectric point 5.2.(2)By conducting the LC-MS / MS mass spectrometric detection with the enzymolysis of the distinct protein in the serum protein between the two groups of subjects,the study proves that the protein spot detected in this research is human haptoglobin(Hp,data code: gi| 1620396 | enb |CAA25927.1),with 348 amino-acid residue and its isoelectric point 6.41.After comparing with the database,the study also shows that there are 5consistent peptide segments in the amino acid sequence in human Hp,with a matching ratio of 16.95%.(3)The results of immunoblotting and ELISA quantitative analysis indicates that,compared with healthy women,the color-developing of the serum in the patients with ovarian cancer is more obvious,thus having high specificity.A quantitative comparison also shows that the content of haptoglobin in patients with ovarian caner is 2.36±1.14(mg/ml),while that of the healthy women is only 1.17 ± 0.32(mg/ml).Its great difference(P < 0.05)offers significance in statistics.(4)Based on the lectin affinity chromatography as well as immunoimprinting,the author finds that the color-developing of FHp(fucosylation haptoglobin)of the patients with ovarian cancer is deeper compared with that of healthy women,thus having relatively high specificity.A semiquantitative analysis after the protein dot blot scanningalso shows that the serum FHp of the patients with ovarian cancer reaches160.80 ±34.87(%),whereas that of the healthy women is only 39.33±4.85(%).Its great difference(P < 0.05)again offers significance in statistics.Conclusion:(1)Haptoglobin differentiates the serum protein between patients with ovarian cancer and healthy women,as has been shown in the two-dimensional gel electrophoresis.(2)The molecular level of Hp of the patients with ovarian cancer is higher than that of the healthy women,as has been proved in immunoimprinting and the quantitative analysis of an ELISA test.(3)The serum FHp of the patients with ovarian cancer is higher than that of the healthy women,as is confirmed by the differential proteome in the lectin affinity chromatography and immunoimprinting.(4)Together with other tumor markers,Hp and FHp can be listed as reference indexes as well as specific indicators when diagnosing patients with early-stage ovarian caner. |
PDF Full Text Request