| ObjectiveTo establish a cytological or histological diagnosis in a patient with suspected lung cancer, flexible bronchoscopy is an essential step in the workup. In the early times, the atpia of the cancer cells are not very obvious or the number is low the sensitivity of purely diagnosis based on cytology is very low. It is due to the fact that there are lots of mucus, blood, inflammatory cells and necrotic material in the slides, resulting in a low detection rate. LCT is successfully used in cervical cytology for the preparation of clear cell monolayers through the removal of mucus, necrotic material and inflammatory cells.The implementation of cervical cancer screening programs has greatly reduced disease incidence and mortality. However a single cytological evaluation remains relatively insensitive, hence need for frequent follow-up investigations. Although the sensitivity of human papillomavinus (HPV) is considered the most significant factor in initiating cervical carcinogenesis, the discordance between the high prevalence of HPV infection and the much lower prevalence of cervical cancer indicates that other genetic events are necessary for progression to malignancy. Establishing a set of scientific and effective diagnostic method is a effective way to improve the positive rate of the diagnosis of lung and cervical cancer. And it is also the key to improve the survival rate and cure rate of the patients with lung and cervical cancer.Our study makes lung and cervical cancer of our major object, and is based on liquid based preparations. We apply FISH on examining the genetic expressing of Hterc gene in the cancer cells. We discuss the correlation between FISH results and cytological and histological diagnosis and evaluate the clinic value of applying on the diagnosis of lung and cervical cancer.MethodsThe present study was performed between January and October 2008 at The First Affiliated Hospital, China Medical University. Two hundred and four were diagnosed withlungcancerbased on histopathology of bronchial biopsy and surgical resection of th e tumors. These 204 patients with lung cancer, as well as 40 randomly selected patients without lung cancer. Histological examination confirmed that there were 104 cases of squamous cell carcinoma (SCC),44 cases of adenocarcinoma (AC), and 56 cases of small cell lung carcinoma (SCLC).102 cases were used for FISH analysis.From March to October 2008,103 cervical liquid-based specimens with outpatients and inpatients, as well as 20 randomly selected specimens with health checkup at The First Affiliated Hospital, China Medical University, were included as study subjects and control subjects, respectively.1. Sample preparation for LCT(1) Two smears were stained with Papanicolaou. Residual brushing was transferred to a small bottle with CytoRich liquid. One millilitre of the mucolytic agent was added to the small bottle with CytoRich liquid, incubated at room temperature for 30 min and vortexed for 10s. Additional mucolytic agent was added to the mixture until the mucus was completely lysed.(2) The mixed liquid was then transferred to a 50-ml tube and centrifuged at 2000 g for 10min. The supernatant was removed and the pellet was resuspended in 7.5ml of distilled water. This suspension was vortexed again and centrifuged at 2000 g for 5min. The supernatant was removed and the pellet was vortexed and transferred to the AutoCyte PREP system, in which slides were automatically prepared and stained.2. Detect hTERC gene by FISH(1) Preparation of the slides.(2) Hybridization of probe and sample. (3) Analysis of FISH results.Result1. Of the 204 brushing samples from patients with lung cancer examined by the conventional CPS method, only 118 were found to have cancer cells, duo to obscuring by mucous material, inflammatory cells and necrotic debris. The diagnostic sensitivity was only 57.8%. However, the LPT showed 71.6% diagnostic sensitivity duo to a clearer background and well preserved cell morphology. This difference in diagnostic sensitivity between the two methods was significant (P＜0.05). When the CPS method and the LPT were evaluated in combination, the diagnostic sensitivity for detecting lung cancer was 76.5%, significantly higher than the CPS method alone (P< 0.01).2. Amplification of TERC was significantly associated with histological diagnosis. Patients with lung cancer had significantly higher percentages of cells with amplification of TERC than did patients of nonmalignant disease (p<0.05). Patients with non-small cell lung cancer had significantly higher percentages of cells with amplification of TERC than did patients of SCLC (p<0.05).3. Of the 102 brushing samples from patients with lung cancer examined by the LCT,84 were found to have cancer cells, the diagnostic sensitivity was 82.4%(84/102) and FISH analysis showed 87.3% (89/102) diagnostic sensitivity.There was no significant difference in diagnostic sensitivity between the two methods (P＞0.05). But the difference in diagnostic sensitivity between the two methods in SCC was significant (P＜0.01).4. Amplification of TERC was significantly associated with both cytological and histological diagnoses (p<0.05). Patients with HSIL or SCC cytology diagnoses had significantly higher percentages of cells with amplification of TERC than did patients with LSIL, ASC-US and lesser conditions (P＜0.005). In histological diagnosis, patients with SCC and CINIII had evidently higher percentages of cells with amplification of TERC than did patients with CINII and lesser conditions either (P<0.005).5. Abnormal hybridization pattern was associated with cytological diagnosis (p<0.05). In subgroup,2-3 hybridization pattern had highest percentages, whereas SHP had highest the correlation coefficient (0.54).Conclusion1. The diagnostic sensitivity of LCT is higher that CPS method in diagnosis of SCLC. When the CPS method and the LPT were evaluated in combination, the diagnostic sensitivity for detecting lung cancer was significantly higher than the CPS method alone2. Detecting hTERC in bronchial brushing pecimens could possibly provided information regarding individual lesions, which is of significant clinical value.3. Detecting hTERC in cervical pecimens could possibly provided information regarding individual lesions, which is of significant clinical value.
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