Studies Of Genetic Mutations Of EGFR And K-ras In The Non-small Cell Lung Cancer By Cytologic Specimens And Intraoperative Effusion Or Washing Cytology In The Clinical Application

Objective:1. To study the feasibility of detecting that EGFR and K-ras gene mutations by cytologic specimens in patients with non-small cell lung cancer, and establish the cytologic detecting methods to give the assistance to the individualized treatment of EGFR-TKIs.2. To explore the the correlation between the genetic mutations of EGFR and K-ras and the protein expression of EGFR, Ki-67and the quantity, heteromorphosis and arranged forms of cancer cells.Materials and methods:1. Collecting the remaining specimens after diagnosis of non-small cell lung cancer in the Department of Pathology in the Chinese Academy of Medical Sciences Cancer Institute&Hospital. The quantity of cancer cells in the TCT smears cells was initially counted by two cytopathologists, and the heteromorphosis and arranged forms of caner cells were recorded at the same time.2.40cases were randomly selected in the cases with abundant cancer cells in residual specimens to perform immunocytochemical detection of EGFR and Ki-67simultaneously.3. The exon18-21th of EGFR gene and the codon12,13th of K-ras gene were amplified by the technology of PCR, and the amplified fragments were then sequenced and analyzed.Results:1. There were20cases of unsatisfactory specimens of the165cytologic specimens extracted DNA were abandoned, and the satisfaction rates was87.9%(145/165).Pleural and pericardial effusion had higher satisfaction rates, which were96.6%and80%respectively, and bronchoscope brush had a lower satisfaction rate (64.7%). In addition, the storage time and the quantity of cancer cells had no statistically significant differences with the satisfaction of DNA extraction.2. In the145patients, the EGFR gene mutation rate of male patients was19.4%(14/72), which was significantly lower than the female patients (37.0%,27/73)(P=0.019). The EGFR gene mutation rate of patients whose age were below60was35.6%(26/73), higher than the patients above60years old (20.8%,15/72)(P=0.048).3. The protein expression of EGFR had no correlation with EGFR gene mutation, and the protein expression of Ki-67also had no correlation with EGFR and K-ras gene mutations.4. The EGFR gene mutation of different quantities of cancer cells were counted(5-10%,11-20%,21-30%,31-40%,41-50%,51-60%,61-70%,71-80%和>80%). According to the Cochran-Armitage for trend test, and the result was Z=4.204, P<0.01. Therefore, the quantity of cancer cells had some correlation with the EGFR gene mutation, and there was a trend of increase for the rate of EGFR gene mutation as the quantity of cancer cells grew. When the quantity of cancer cells was above30%, the rate of EGFR gene mutation had an obvious trend of increase.5. When the heteromorphosis of cells was divided into mild, moderate and severe, and the EGFR gene mutation rate was7.7%,27.6%and40.7%respectively. The P-value between the co-exist of the three, mild and non-mild or severe and non-severe and mild and severe were all larger than0.05, therefore, there was no statistically significant difference between the heteromorphosis of cells and the EGFR gene mutation. When the arranged forms of cells was divided into the mainly papilliary, papillary and scattered, and the mainly scattered, and the EGFR gene mutation rate was24.6%,29.2%and34.4%respectively. The P-value between the co-exist of the three, the mainly papillary and non-papillary arrange or the mainly scattered and non-scattered arrange and the mainly papillary and the mainly scattered were all larger than0.05, therefore, there was no statistically significant difference between the arranged forms of cells and the EGFR gene mutation.Conclusions:1. Cytologic specimens were feasible and validated for detecting the EGFR and K-ras gene mutations, especially in patients whose histologic specimens couldn’t be obtained.2. The quantity of cancer cells had some correlation with the EGFR gene mutation, and there was a trend of increase for the rate of EGFR gene mutation as the quantity of cancer cells grew. When the quantity of cancer cells was above30%, the rate of EGFR gene mutation had an obvious trend of increase.3. Both the heteromorphosis and the arranged forms of cancer cells had no correlation with the EGFR gene mutation. Therefore, the heteromorphosis and the arranged forms of cancer cells couldn’t be used as predictors of the EGFR gene mutation. Abstract:Objective To evaluate the clinical application value of intraoperative effusion or washing cytology and its accuracy. Methods521patients in our hospital were enrolled. Their cytological diagnosis and histological diagnosis after surgery were compared to analyses the characteristics of the smears by reviewing the false-positive and false-negative slides and to evaluate its accuracy and clinical application value. Results During the521intraoperatively obtained samples the detection rate of the malignant cells was40.32%, whereas the detection rate of the cases that had peritoneal metastasis by histopathology was84.9%. In the current study, there were9false-positive and18false-negative cases. The main reason for the false-positive was that it’s difficult to discriminate the endometriosis or the mesothelial cell hyperplasia from the well-differentiated adenocarcinoma, while the main reason for the false-negative was the low detection sensitivity caused by the Inflammatory cells and the hemocytes. Conclusion The rate of detection of intraoperative cytology were relatively high and it had certain accuracy. Therefore it had reference value for the patients’ follow-up treatments and prognosis. Abstract:Objective To explore the influencing factor of cytological detection ration of intraoperative ascites or washing in low malignant potential borderline cystadenoma of ovary and its’cells’morphological characteristics in the cytological slides. Methods521patients in our hospital were enrolled. Their cytological diagnosis and histological diagnosis after surgery were followed up to explore the influencing factor of cytological detection ration of intraoperative ascites or washing in low malignant potential borderline cystadenoma of ovary. Reading the cytological slides of low malignant potential borderline cystadenoma of ovary and observing the characteristics to explore the morphological traits and the value of diagnosis. Results30low malignant potential borderline cystadenomas of ovary were found of the521intraoperatively obtained samples.16of them were detected, accounting for53.3%. Serous low malignant potential borderline cystadenomas of ovary were much easier to be detected compared to the mucous(P=0.046).9borderline tumors which had no implants were positive in cytology, thus cytology could find tiny implants which may be neglected by naked eyes or pathology. Cells’ heteromorphosis of low malignant potential borderline cystadenoma of ovary was unconspicuous, but the epithelial structure could point out tumors not mesothelial cells. Hyperplastic mesothelial cells which had papillary fragment could lead to cytological misdiagnose, but the papillary fragment was comparatively tiled and didn’t have third dimension. The most significances in differential diagnosis of morphological traits between low malignant potential borderline tumors and adenocarcinoma were the obviously enlargement of chromatin and the irregularity of karyotype. Conclusion The cytological detection ration of the serous low malignant potential borderline cystadenoma of ovary was comparatively higher. In the cytological slides of intraoperative ascites or washing, cells of the low malignant potential borderline cystadenoma or the adenocarcinoma of ovary were comparatively easier to distinguish with hyperplastic mesothelial cells, while cells of low malignant potential borderline cystadenoma were more difficult to be distinguished from the adenocarcinoma cells and the most significances in differential diagnosis were the obviously enlargement of chromatin and the irregularity of karyotype.