The Research Of Liquid-based Cytology Test In Combination With P16～(INK4a) And Cyclin E In Early Diagnosis Of Cervical Process

Objective To investigate the value of liquid-based cytology test(LCT)in combination with P16^INK4a and cyclin E in the early diagnosis ofcervical intraepithelial neoplasia(CIN).Methods 225 patients were investigated in the present study,including 56 squamous cell carcinomas(SCC), 41 high grade squamousintraepithelial lesions (HSIL), 76 low grade squamous intraepitheliallesions(LSIL), 32 atypical squamous cell lesions(ASC) and 20 caseswithin normal limits(WNL). LCT was used in all the patients.Immunohistochemistry was performed to examine the expressions ofP16^INK4a and cyclin E. The pathological results were used as the goldstandard to evaluate the results of traditional Pap smear and LCT method,LCT method and the combination of LCT with P16^INK4a and cyclin E; theexpression of P16^INK4a and cyclin E in LCT and paraffinum histologicalsection.Results1. Compared with the traditional Pap smear and LCT method, theunsatisfactory ratio of conventional Pap smear is 12.44ï¼…(28/225), butLCT smears shows content resulting in 225 cases. The process diagnosecoincidence of conventional Pap smear is LSIL49.38ï¼…(40/81), HSIL56.10ï¼…(23/41), SCC82.14ï¼…(46/56), respectively. The coincidenceof LCT smears with biopsy diagnose are LSIL52.63ï¼…(40/76),HSIL68.29ï¼…(28/41), SCC 87.5ï¼…(49/56), which show great apparentdifference in HSIL and SCC between them(x²=7.635, x²=8.157, P＜0.05).And no significance difference of LSIL(x²=3.142, P＞0.05). Depend onpathologic diagnosis of ASC' biopsy and operation exemplar, thecellule diagnose of traditional Pap smear suggest there are 17 subjects oftrails including normal or chronic inflammation 13, 1 for SCC, 1 forCINâ… , 2 for CINâ…¡-â…¢. And in LCT smears trails is 27 of 32 for normalor chronic inflammation, 2 for CINâ… , 2 for CINâ…¡-â…¢, 1 for SCC. Thereis no significance difference of statistic meaning (x²=1.851, P＞0.05)Thus we can draw a conclusion that LCT smears can boost the diagnoseratio of HSIL, while SCC fails to boost diagnose ratio of LSIL and ASC.2. Expression of P16^INK4a and cyclin E in process of the cervix.2.1 Expression of P16^INK4a and comparison the consequence of LCTand LCT in combination with P16^INK4a. In all cases, normal cervix cellulaepithelialis free of coloration and cervix cellula epithelial glands alsoshow negative-chromatosis of P16^INK4a. The accuracies of LCT incombination with P16^INK4a were 89.29ï¼…(50/56), 85.71ï¼…(30/35),68.75ï¼…(33/48), 12.55ï¼…(4/32) in 56 SCC, 35 HSIL, 48 LSIL, 32 ASCrespectively. Expression of P16^INK4a shows strength masculine and it hasclose-up relation ship with SIL's classification. With full-thickness the rise of SIL degrade, by which enhance the P16^INK4a appearance and filledwith full-thickness. Between LSIL and HSIL, there are obviouslydifference (x²=15.431,P＜0.05). Among MNL, SIL and SCC, there areobviously difference(x²=33.637, x²=18.257, P<0.05). Compared withLCT method and LCT in combination with P16^INK4a: there are obviouslydifference between them in the class LSIL, HSIL,P=0.002, P=0.043,respectively, there is no diference in SCC, P＞0.05.2.2 Expression of cyclin E and comparison the consequence of LCTand LCT in combination with cyclin E. In all cases, normal cervix cellulaepithelialis free of coloration and cervix cellula epithelial glands alsoshow negative-chromatosis of cyclin E. The accuracies of LCT incombination with cyclin E were 85.71ï¼…(48/56),74.29ï¼…(26/35), 83.33ï¼…(40/48), 9.38ï¼…(3/32) in 56 SCC, 35 HSIL, 48 LSIL, 32 ASC respectively.Expression of cyclin E show high masculine in LSIL.but there isdifference with P16^INK4a. Between LSIL and HSIL there are no obviouslydifference (x²=2.271,P＞0.05). The identical of P16^INK4a and cyclin E is:heavier the process, stronger the masculine expression. Among MNL,SIL and SCC, there are obviously difference (x²=15.487,x²=17.382,P＜0.05). Compared with LCT method and LCT incombination with cyclin E, there is no diference between them in theclass of HSIL and SCC, P=0.5; P＞0.05 respectively. There are obviouslydifference between them in the class LSIL, P=0.000. LCT in combination with cyclin E boost the diagnose ratio of LSIL obviously.3. Comparison the consequence of LCT and LCT in combination withP16^INK4a and cyclin E. Compared with the pathological results, theaccuracies of LCT method were 52.63ï¼…, 68.29ï¼…, 87.5ï¼…in LSIL, HSIL,SCC respectively. The accuracies of LCT in combination with P16^INK4aand cyclin E were 87.50ï¼…, 88.57ï¼…, 91.07ï¼…in LSIL, HSIL and SCCrespectively. HSIL, P=0.016; LSIL, P=0.000; The difference wassignificant, there is no diference in SCC, P＞0.05. The difference wassignificant. In 32 cases of ASC, 3 of all show strong masculine ofP16^INK4a and its pathology diagnose is cancer in situ which involve gland,1 for moderately differentiated SCC, 1 for cancer in situ; among the total,1 for show strong masculine of cyclin E, 1 for masc expression, 1 fornegative; In 2 CINâ… , 1 show masculine of P16^INK4a and cyclin E, 1 shownegative both. As account, the accuracy of LCT in combination withP16^INK4a and cyclin E was 80ï¼…(4/5), compared with LCT method andLCT in combination with P16^INK4a and cyclin E has more value in thediagnose of ASC.4. Comparison of the expression of P16^INK4a and cyclin E in LCT andparaffinum histological section. In 56 SCC, the expression of P16^INK4aand cyclin E in LCT is 89.29ï¼…(50/56) and 85.71ï¼…(48/56) respectively.The expression in paraffinum histological section is 85.71ï¼…(48/56) and80.36ï¼…(45/56), there is no diference between them(x²=2.637, x²=2.815, P ＞0.05).Conclusion1. LCT smears has obviously superiority compare with pap smear. Itnot only provides sufficient sample but also improves the quality ofsmear, which makes it easy to identify the anomal-cell and elevate thediagnose rate of masculine cell. According to the TBS can reduce thesubjective reflection and cut down the false positive rate and falsenegative rate, it has important significance to elevate the sensitively andspecificity of screening.2. The overwxpression of P16^INK4a is concerned with thecarcinogenesis and development of cervical carcinoma. Expression ofP16^INK4a shows strength masculine in HSIL and SCC, and it has close-uprelation ship with SIL's classification. With full-thickness the rise of SILdegrade, by which enhance the P16^INK4a appearance and filled withfull-thickness. Among MNL, SIL and SCC, there are obviouslydifference.3. P16^INK4a is participate for the development of cervical carcinoma incoordination with cyclin E. P16^INK4a has higher diagnose accordance ratein HSIL, cyclin E has higher diagnose accordance rate in LSIL. Theyhave some resemblance in control mechanism and pathway, they bothparticipate the cell cycle control.4. P16^INK4a can act as a new target in uterine cervix cancer'oncogene therapy, for its high expression both in CIN and SCC.5. LCT in combination with P16^INK4a and cyclin E may be a usefulmethod in screening of CIN, especially in the diagnosis of ASC. It hashigh pathologic diagnosis accordance rate than purely LCT, thus canboost diagnose ratio of uterine cervix cancer and precancerous process. Inaddition, compared with histological specimen, LCT in combination withP16^INK4a and cyclin E have advantage of draw materiale convenience andpatient little pain, so the new method is worth to be spread.