ALEX1 Expression In Cervical Cancer Tissues And Effect Biological Characteristics Of On Cervical Cancer Cell

Cervical cancer is a serious disease that affects women’s physical and mental health. The World Health Organization(WHO) reported that,cervical cancer comprises 12 % of all cancers globally and it is second most common gynecological malignancy after breast cancer in the world.ALEX1 is a novel subgroup within the armadillo family which has two Arm repeats domains and contain at protein N-terminal has a hydrophobic transmembrane region. ALEX1 are widely expressed in normal tissues except for liver and thymus tissue with low expression and no expression in peripheral blood leukocytes. And ALEX1 low or no expression in tumor tissue from Epithelial tissue. The present study showed the ALEX1 protein is associated with tumorigenesis and overexpression of ALEX1 suppresses colony formation of human carcinoma cells. But the has not been reported that ALEX1 is associated with tumorigenesis in cervical cancer. This paper will explore the expression of ALEX1 in cervical cancer, and study its effect on the biological characteristics of on cervical cancer cell.Part 1: ALEX1 expression in cervical cancer tissues and clinical significanceObjective: To study the expression of ALEX1 protein in cervical cancer tissues and adjacent tissues; and to analyze the difference of A expression and its clinical significance.Methods: Collection of 53 clinical cervical cancer and adjacent tissues;fixed with poly formaldehyde and dehydration with ethanol; paraffin embedded and sliced. Evaluate the ALEX1 protein expression differences in clinical cervical cancer and adjacent tissues using immunohistochemical.And analyzed the clinical characteristics effect ALEX1 protein expression in tissues.Result: The results show that ALEX1 protein expression is high in 44 of 53(83.0 %) cervical carcinoma tissues and in 13 of 53(24.5 %)non-cancerous tissues(*P ﹤ 0.05). ALEX1 was mainly expressed in the cytoplasm of cancer tissues, but adjacent tissues showed mainly expression in the nuclear.Conclusion: ALEX1 protein expression is significantly increased in cervical cancer tissues compared with adjacent tissues. ALEX1 was mainly expressed in the cytoplasm of cancer tissues, but adjacent tissues showed mainly expression in the nuclear.Part 2: Effects of si RNA silencing of the ALEX1 on cell cycle,cell proliferation,and apoptosis in cervical cancer cellsObjective: Designed and synthesized three small interfering RNA targeting ALEX1 gene was transfected into Hela and selected the best sequence. To investigate the effect of silencing ALEX1 on the biological characteristics of cervical cancer cells Hela.Methods: Designed and synthesized three small interfering RNA targeting ALEX1 gene was transfected into Hela and selected the best sequence. Evaluated the effect of silencing ALEX1 on Hela cells cycle and apoptosis was analysied by flow cytometry. And effect of ALEX1 interference on Hela cells proliferation was analysied by CCK-8 assay.Result: QRT-PCR and Western blotting indicated that the RNAi-3 was the best for cervical cancer Hela cells. Flow cytometry showed that cells in S phase rate of Hela cells transfected with RNAi-3(30.00 ± 1.77 %) was significantly higher than untreated group(16.53 ± 0.93 %) and negative control group(14.41±0.86 %)(*P﹤0.05); and apoptosis rate of Hela cells transfected with RNAi-3(16.41 ± 0.59 %) was significantly higher than untreated group(2.31±0.23 %) and negative control group(3.15±0.33 %)(*P ﹤ 0.05). Proliferation of cells Hela transfected with RNAi-3 was inhibited compared with control group and blank group(*P﹤0.05).Conclusion: Silencing the expression of ALEX1 can promote the apoptosis and block the cell cycle progression of human cervical cancer cells Hela; and proliferation of Hela cells was inhibited.Part 3: Silencing ALEX1 enhances Res’ s ability to kill cervical cancerObjective: To study the effects of ALEX1 silence to Res on cell rate of survival and apoptosis; and the changes of IC50 were determined after Res treatment at different time.Methods: Effects of ALEX1 silence to sensitivity of Res in cervical cancer cells was analysied by CCK-8; Concentration setting: 0、50、100、200、400、600、800 μmol· L-1.Result: This study showed that cells treated with Res and transfected with RNAi-3 growth was inhibited than untreated group and negative control group. And 48,72 h after treated with Res, IC50 was significantly reduced than untreated group and negative control group. Apoptosis rate of Hela cells transfected with RNAi-3 and treated with Res(36.33±3.17 %)was significantly higher than negative group(16.58 ± 1.95 %) and untreated control group(12.28±1.28 %))(*P﹤0.05).Conclusion: In 48、72 h, silencing ALEX1 enhances Res’ s ability to kill cervical cancer, and the sensitivity of Res to cervical cancer was increased. Silencing ALEX1 expression, enhanced Res to promote the occurrence of apoptosis of Hela cells.