LRIG3在宫颈鳞癌组织中的表达及体内外干扰LRIG3表达的实验研究
发布时间：2018-07-29 08:31
【摘要】：背景 宫颈癌是全世界范围内同时也是我国最常见的恶性肿瘤之一，其发生率位居女性生殖系统恶性肿瘤第一位，且有逐年年轻化的趋向。近年来，随着手术切除、化学治疗、放射治疗、及其它综合性治疗措施的应用，对宫颈癌的治疗有了一定程度的进展，但疗效并不十分理想。宫颈癌的病因及发生机制目前尚不十分明了，许多研究证明其发生、发展和浸润、转移是多因素、多阶段、多基因共同参与的非常复杂的过程，因此，探讨宫颈癌的发病机制，寻找更有效的治疗措施是当前研究的热点课题之一。 LRIGs是最近发现的一组肿瘤相关基因，该家族共有三个成员：LRIG1、LRIG2、LRIG3。LRIGs编码一组结构上较为相似的膜整合蛋白，蛋白分为胞外段、跨膜区和一个胞浆内尾巴，胞外段蛋白含有富含亮氨酸重复序列（LRRs）和3个免疫球蛋白样结构域。研究表明LRIG的表达能够对EGFR信号通路进行调控，从而起到抑制肿瘤生长的作用，但是我们对其分子和信号机制尚不清楚。近年来许多研究表明，，LRIGs基因家族在垂体瘤、神经胶质瘤和皮肤鳞状细胞癌等多种肿瘤中具有抑癌作用。 LRIG3定位于人类基因组12q13，在人类肿瘤中LRIG3是高表达还是低表达尚不十分清楚，对于其在宫颈癌中表达的相关研究较少，对于其在宫颈癌细胞株中发挥的生物学作用尚未见报道。LRIG3和宫颈癌的发生发展是否有相关性，抑制LRIG3表达对宫颈鳞癌的影响是本研究需要探讨的核心。 目的 检测宫颈鳞癌组织、宫颈上皮内瘤变组织和正常宫颈粘膜组织中LRIG3、EGFR和Ki67的表达，并分析其与宫颈鳞癌浸润转移的关系。观察LRIG3反义核苷酸转染到宫颈癌Hela229细胞中后对LRIG3和EGFR的表达以及对细胞增殖、转移以及细胞周期的影响。观察LRIG3反义核苷酸对宫颈癌裸鼠移植瘤的影响。 第一部分LRIG3在宫颈鳞癌组织中的表达及意义 方法 应用免疫组织化学和原位杂交检测45例宫颈鳞癌组织、20例CIN组织和30例正常宫颈黏膜组织中LRIG3、EGFR和Ki67表达，并结合临床病理资料对结果进行分析。 结果 1. LRIG3蛋白和mRNA在宫颈鳞状细胞癌组织中的阳性表达率分别为35.56%和48.89%，显著低于CIN组织（分别为70%、80%）和正常宫颈粘膜组织（皆为100%），两两间比较差异均有统计学意义(P0.05)。其蛋白和mRNA的表达强度与/和患者的年龄以及肿瘤大小无关（P均0.05），而与肿瘤的浸润程度、临床分期以及淋巴结转移成负相关（P均0.05）。 2. EGFR蛋白和mRNA在宫颈鳞状细胞癌组织中的阳性表达率分别为80%和84.44%，显著高于CIN组织（分别为35%、45%）和正常宫颈粘膜组织（分别为6.67%、16.67%），两两间比较差异均有统计学意义(P0.05)。在宫颈鳞状细胞癌组织中EGFR蛋白和mRNA的表达强度与患者的年龄、肿瘤大小、临床分期无关（P均0.05），而与肿瘤的浸润程度以及淋巴结转移成正相关（P均0.05）。 3. Ki67蛋白和mRNA在宫颈鳞状细胞癌组织中的阳性表达率分别为86.67%和91.11%，显著高于CIN组织（分别为30%和50%）和正常宫颈粘膜组织（分别为3.33%、23.33%），两两间比较差异均有统计学意义(P0.05)。Ki-67的表达与宫颈鳞癌各临床病理因素均无相关性((P0.05)。 4. LRIG3和EGFR在宫颈鳞状细胞癌组织中的表达呈负向相关, LRIG3和Ki67在宫颈鳞状细胞癌组织中的表达亦呈负向相关。 第二部分LRIG3的表达调控对宫颈鳞癌Hela229细胞周期、凋亡、侵袭力的影响 方法 1.设计合成LRIG3反义寡核苷酸（LRIG3ASODN）、正义寡核苷酸序列（LRIG3SODN）以及错义寡核苷酸序列（LRIG3MSODN），用脂质体Lipofectamine2000包裹转染Hela229细胞，作为转染组细胞。将脂质体转染的细胞作为空白对照组细胞。 2. Western blot检测不同浓度(150μg/ml、200μg/ml、250μg/ml) LRIG3ASODN、LRIG3MSODN和LRIG3SODN转染细胞24h、48h及72h后LRIG3和EGFR蛋白表达情况。 3. RT-PCR检测不同浓度(150μg/ml、200μg/ml、250μg/ml) LRIG3ASODN、LRIG3MSODN和LRIG3SODN转染细胞24h、48h及72h后LRIG3和EGFRmRNA表达情况。 4.应用MTT检测细胞增殖情况，细胞流式法测定细胞凋亡，细胞基质粘附实验检测细胞粘附能力，Transwell小室法检测细胞的侵袭能力。 结果 1. ASODN组中LRIG3蛋白和mRNA表达较SODN组、MSODN组、空白对照组明显降低（P0.05），且有浓度依赖性和时间依赖性，LRIG3的表达随转染浓度的增加以及时间的延长而减少。 2. ASODN组中EGFR蛋白和mRNA表达较SODN组、MSODN组、空白对照组明显增加（P0.05），随转染浓度的增加及时间的延长EGFR的表达随之升高。EGFR表达和LRIG3表达呈负相关。 3. MTT实验表明LRIG3ASODN组Hela229细胞增殖率较其它组别明显升高（P0.05），随着LRIG3ASODN转染浓度增加，表达抑制时间延长，细胞增殖率明显升高，差异有统计学意义（P0.05）。 4.转染LRIG3ASODN后Hela229细胞凋亡率明显降低，随着转染浓度的增加，时间延长，细胞的凋亡率随之降低。 5.细胞基质粘附实验表明转染LRIG3ASODN后使Hela229细胞粘附率明显增高，随着转染浓度的增加，时间延长，细胞的粘附率随之升高。 6. Transwell实验表明转染LRIG3ASODN后使得Hela229细胞的迁徙能力明显增高，ASODN组穿膜细胞数（246±10）明显高于SODN组（168±10）、MSODN组（177±8）和空白对照组（176±6）。 第三部分LRIG3表达调控对人宫颈鳞癌裸鼠移植瘤生长的影响 方法 将1×107个Hela229细胞注射到裸鼠皮下构建宫颈鳞癌裸鼠移植瘤模型，将LRIG3ASODN皮下注射到瘤体内后观察裸鼠移植瘤的生长状态。测量各组种植瘤瘤体体积。Western blot和RT-PCR检测裸鼠移植瘤中LRIG3和EGFR蛋白和mRNA表达。 结果 1. ASODN LRIG3组注射后18-30天裸鼠细胞瘤体积较其它两组明显增高（P0.05）。 2. Western blot和PT-PCR检测裸鼠移植瘤中LRIG3蛋白和mRNA表达，显示ASODN组LRIG3表达较MSODN和空白对照组明显降低（P0.05）。 3. Western blot和PT-PCR检测裸鼠移植瘤中EGFR蛋白和mRNA表达，显示ASODN组EGFR表达较MSODN和空白对照组明显升高（P0.05）。 结论 1.宫颈鳞癌组织和CIN组织中LRIG3呈低表达，EGFR高表达，其表达量与宫颈鳞癌的浸润和转移密切相关。提示LRIG3通过对EGFR的负反馈调节起到抑制宫颈鳞癌发生发展的作用。 2. LRIG3ASODN能显著下调宫颈鳞癌Hela229细胞中LRIG3蛋白和mRNA的表达，同时EGFR蛋白和mRNA表达上升，能抑制宫颈鳞癌细胞凋亡，增强细胞侵袭能力。提示LRIG3在宫颈鳞癌细胞增殖、凋亡及侵袭力中起着重要作用。 3. LRIG3ASODN可导致宫颈鳞癌裸鼠成瘤速度加快，体积增加，同时移植瘤组织中LRIG3蛋白和mRNA的表达显著降低，EGFR蛋白和mRNA表达增高。
[Abstract]:background
Cervical cancer is one of the most common malignant tumors in the world and is one of the most common malignant tumors in China. Its incidence is the first malignant tumor in female reproductive system, and it has a tendency to be young year by year. In recent years, with the application of surgical resection, chemical therapy, radiation therapy, and other comprehensive treatment measures, the treatment of cervical cancer has a certain course. The etiology and mechanism of cervical cancer are still not very clear. Many studies have proved that the occurrence, development and infiltration of cervical cancer are a very complex process involving multiple factors, multi stages and multiple genes. Therefore, the study of the pathogenesis of cervical cancer and the search for more effective treatment measures are currently studied. One of the hot topics in the study.
LRIGs is a recently discovered group of tumor related genes. There are three members of the family: LRIG1, LRIG2, and LRIG3.LRIGs, which encode a group of relatively similar membrane integrins. The proteins are divided into extracellular, transmembrane and cytoplasmic tails, and exosin contains high leucine rich repeat (LRRs) and 3 immunoglobulin like domains. Studies have shown that the expression of LRIG can regulate the EGFR signaling pathway, thus inhibiting the growth of the tumor, but we are not clear about its molecular and signal mechanisms. In recent years, many studies have shown that the LRIGs gene family has a tumor suppressor effect in a variety of tumors, such as pituitary adenoma, glioma and skin squamous cell carcinoma.
LRIG3 is located in the human genome 12q13. The high expression or low expression of LRIG3 in human tumor is not very clear. There are few related studies on its expression in cervical cancer. There is no report on the correlation between the development of.LRIG3 and cervical cancer and the inhibition of the expression of LRIG3 in cervical cancer cell lines. The impact on cervical squamous cell carcinoma is the core of this study.
objective
To detect the expression of LRIG3, EGFR and Ki67 in cervical squamous cell carcinoma, cervical intraepithelial neoplasia and normal cervical mucosa, and to analyze the relationship with invasion and metastasis of squamous cell carcinoma of the cervix. The expression of LRIG3 antisense nucleotides transfected to Hela229 cells of cervical cancer and the expression of LRIG3 and EGFR and the shadow of cell proliferation, metastasis and cell cycle were observed. To observe the effect of LRIG3 antisense nucleotide on cervical cancer xenografts in nude mice.
Expression and significance of LRIG3 in cervical squamous cell carcinoma
Method
Immunohistochemistry and in situ hybridization were used to detect the expression of LRIG3, EGFR and Ki67 in 45 cases of cervical squamous cell carcinoma, 20 CIN tissues and 30 normal cervical mucosa tissues, and the results were analyzed with the clinicopathological data.
Result
The positive expression rates of 1. LRIG3 protein and mRNA in cervical squamous cell carcinoma were 35.56% and 48.89%, respectively, significantly lower than that of CIN tissues (70%, 80%, respectively) and normal cervical mucosa (100%). The 22 differences were statistically significant (P0.05). The expression intensity of protein and mRNA and the age of the patients and the size of the tumor None (P = 0.05), but negatively correlated with tumor infiltration, clinical stage and lymph node metastasis (P = 0.05).
The positive expression rates of 2. EGFR protein and mRNA in cervical squamous cell carcinoma were 80% and 84.44% respectively, which were significantly higher than those of CIN (35%, 45%) and normal cervical mucosa (6.67%, 16.67%, respectively, 6.67%, 16.67%, respectively). The expression of EGFR protein and mRNA in the squamous cell carcinoma of the cervix was significant. No correlation was found with age, tumor size and clinical stage (P = 0.05), but positively correlated with tumor infiltration and lymph node metastasis (P 0.05).
The positive expression rates of 3. Ki67 protein and mRNA in cervical squamous cell carcinoma were 86.67% and 91.11% respectively, which were significantly higher than those of CIN tissues (30% and 50%) and normal cervical mucosa (3.33%, 23.33%, respectively). There were significant differences in 22 differences (P0.05).Ki-67 expression was not different from the clinical and pathological factors of cervical squamous cell carcinoma. P0.05.
4. the expression of LRIG3 and EGFR in cervical squamous cell carcinoma was negatively correlated, and the expression of LRIG3 and Ki67 in cervical squamous cell carcinoma was also negatively correlated.
The second part is the effect of LRIG3 expression regulation on cell cycle, apoptosis and invasiveness of cervical squamous cell carcinoma Hela229.
Method
1. the LRIG3 antisense oligonucleotide (LRIG3ASODN), the just oligonucleotide sequence (LRIG3SODN) and the missense oligonucleotide sequence (LRIG3MSODN) were designed and synthesized. The transfected Hela229 cells were transfected with liposome Lipofectamine2000 as the transfected cells. The cells transfected with liposomes were used as blank control cells.
The expression of 24h, 48h and 72h after transfection of 24h, 48h and 72h was detected by 2. Western blot (150 g/ml, 200, g/ml, 250 mu g/ml) LRIG3ASODN, LRIG3MSODN and LRIG3SODN.
3. RT-PCR was used to detect the expression of 24h and 48h and 72h after transfection at different concentrations (150 g/ml, 200 g/ml, 250 g/ml) LRIG3ASODN, LRIG3MSODN and LRIG3SODN.
4. the cell proliferation was detected by MTT, cell apoptosis was measured by flow cytometry, cell adhesion test was used to detect cell adhesion ability, and cell invasion ability was detected by Transwell chamber method.
Result
The expression of LRIG3 protein and mRNA in group 1. ASODN was significantly lower than that in group SODN, in group MSODN and in blank control group (P0.05), and there was a concentration dependence and time dependence. The expression of LRIG3 decreased with the increase of transfection concentration and the prolongation of time.
The expression of EGFR protein and mRNA in group 2. ASODN was significantly higher than that in group SODN, and in MSODN group, the blank control group was significantly increased (P0.05). The expression of.EGFR and LRIG3 expressed a negative correlation with the increase of the transfection concentration and the prolongation of the expression of EGFR.
3. MTT test showed that the proliferation rate of Hela229 cells in group LRIG3ASODN was significantly higher than that of other groups (P0.05). With the increase of LRIG3ASODN transfection concentration, the expression inhibition time was prolonged and the cell proliferation rate was significantly increased, the difference was statistically significant (P0.05).
4. the apoptosis rate of Hela229 cells decreased significantly after transfection of LRIG3ASODN. With the increase of transfection concentration and the prolongation of time, the apoptosis rate of cells decreased.
The adhesion test of 5. cell matrix showed that the adhesion rate of Hela229 cells increased obviously after transfection of LRIG3ASODN. With the increase of transfection concentration, the time was prolonged and the adhesion rate of cells increased.
6. Transwell test showed that the migration ability of Hela229 cells increased significantly after transfection of LRIG3ASODN. The number of membrane cells in ASODN group (246 + 10) was significantly higher than that in group SODN (168 + 10), MSODN group (177 + 8) and blank control group (176 + 6).
The third part is the effect of LRIG3 expression regulation on the growth of human cervical squamous cell carcinoma xenografts in nude mice.
Method
1 x 107 Hela229 cells were injected into nude mice to construct nude mice model of nude mice, and the growth state of nude mice was observed after subcutaneous injection of LRIG3ASODN into the tumor. The expression of LRIG3 and EGFR protein and mRNA in the transplanted tumor of nude mice was measured by.Western blot and RT-PCR.
Result
In 1. ASODN LRIG3 group, the tumor volume in nude mice was significantly higher than that in the other two groups on the 18-30 day after injection (P0.05).
2. Western blot and PT-PCR detected the expression of LRIG3 protein and mRNA in nude mice xenograft. The expression of LRIG3 in ASODN group was significantly lower than that in MSODN and blank control group (P0.05).
3. Western blot and PT-PCR detected the expression of EGFR protein and mRNA in nude mice xenograft. The expression of EGFR in ASODN group was significantly higher than that in MSODN and blank control group (P0.05).
conclusion
1. the expression of LRIG3 in cervical squamous cell carcinoma tissues and CIN tissues is low, and the expression of EGFR is highly expressed. The expression of LRIG3 is closely related to the invasion and metastasis of cervical squamous cell carcinoma. It suggests that the negative feedback regulation of EGFR plays a role in inhibiting the development of cervical squamous cell carcinoma.
2. LRIG3ASODN can significantly reduce the expression of LRIG3 protein and mRNA in the Hela229 cells of cervical squamous cell carcinoma, and increase the expression of EGFR and mRNA, which can inhibit the apoptosis of cervical squamous cell carcinoma cells and enhance the invasive ability of the cells, suggesting that LRIG3 plays an important role in the proliferation, apoptosis and invasion of cervical squamous cell carcinoma cells.
3. LRIG3ASODN could lead to accelerated tumor formation and increased volume of cervical squamous cell carcinoma in nude mice. The expression of LRIG3 protein and mRNA in the transplanted tumor tissue was significantly decreased, and the expression of EGFR protein and mRNA was increased.
【学位授予单位】：郑州大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R737.33
本文编号：2152099
[Abstract]:background
Cervical cancer is one of the most common malignant tumors in the world and is one of the most common malignant tumors in China. Its incidence is the first malignant tumor in female reproductive system, and it has a tendency to be young year by year. In recent years, with the application of surgical resection, chemical therapy, radiation therapy, and other comprehensive treatment measures, the treatment of cervical cancer has a certain course. The etiology and mechanism of cervical cancer are still not very clear. Many studies have proved that the occurrence, development and infiltration of cervical cancer are a very complex process involving multiple factors, multi stages and multiple genes. Therefore, the study of the pathogenesis of cervical cancer and the search for more effective treatment measures are currently studied. One of the hot topics in the study.
LRIGs is a recently discovered group of tumor related genes. There are three members of the family: LRIG1, LRIG2, and LRIG3.LRIGs, which encode a group of relatively similar membrane integrins. The proteins are divided into extracellular, transmembrane and cytoplasmic tails, and exosin contains high leucine rich repeat (LRRs) and 3 immunoglobulin like domains. Studies have shown that the expression of LRIG can regulate the EGFR signaling pathway, thus inhibiting the growth of the tumor, but we are not clear about its molecular and signal mechanisms. In recent years, many studies have shown that the LRIGs gene family has a tumor suppressor effect in a variety of tumors, such as pituitary adenoma, glioma and skin squamous cell carcinoma.
LRIG3 is located in the human genome 12q13. The high expression or low expression of LRIG3 in human tumor is not very clear. There are few related studies on its expression in cervical cancer. There is no report on the correlation between the development of.LRIG3 and cervical cancer and the inhibition of the expression of LRIG3 in cervical cancer cell lines. The impact on cervical squamous cell carcinoma is the core of this study.
objective
To detect the expression of LRIG3, EGFR and Ki67 in cervical squamous cell carcinoma, cervical intraepithelial neoplasia and normal cervical mucosa, and to analyze the relationship with invasion and metastasis of squamous cell carcinoma of the cervix. The expression of LRIG3 antisense nucleotides transfected to Hela229 cells of cervical cancer and the expression of LRIG3 and EGFR and the shadow of cell proliferation, metastasis and cell cycle were observed. To observe the effect of LRIG3 antisense nucleotide on cervical cancer xenografts in nude mice.
Expression and significance of LRIG3 in cervical squamous cell carcinoma
Method
Immunohistochemistry and in situ hybridization were used to detect the expression of LRIG3, EGFR and Ki67 in 45 cases of cervical squamous cell carcinoma, 20 CIN tissues and 30 normal cervical mucosa tissues, and the results were analyzed with the clinicopathological data.
Result
The positive expression rates of 1. LRIG3 protein and mRNA in cervical squamous cell carcinoma were 35.56% and 48.89%, respectively, significantly lower than that of CIN tissues (70%, 80%, respectively) and normal cervical mucosa (100%). The 22 differences were statistically significant (P0.05). The expression intensity of protein and mRNA and the age of the patients and the size of the tumor None (P = 0.05), but negatively correlated with tumor infiltration, clinical stage and lymph node metastasis (P = 0.05).
The positive expression rates of 2. EGFR protein and mRNA in cervical squamous cell carcinoma were 80% and 84.44% respectively, which were significantly higher than those of CIN (35%, 45%) and normal cervical mucosa (6.67%, 16.67%, respectively, 6.67%, 16.67%, respectively). The expression of EGFR protein and mRNA in the squamous cell carcinoma of the cervix was significant. No correlation was found with age, tumor size and clinical stage (P = 0.05), but positively correlated with tumor infiltration and lymph node metastasis (P 0.05).
The positive expression rates of 3. Ki67 protein and mRNA in cervical squamous cell carcinoma were 86.67% and 91.11% respectively, which were significantly higher than those of CIN tissues (30% and 50%) and normal cervical mucosa (3.33%, 23.33%, respectively). There were significant differences in 22 differences (P0.05).Ki-67 expression was not different from the clinical and pathological factors of cervical squamous cell carcinoma. P0.05.
4. the expression of LRIG3 and EGFR in cervical squamous cell carcinoma was negatively correlated, and the expression of LRIG3 and Ki67 in cervical squamous cell carcinoma was also negatively correlated.
The second part is the effect of LRIG3 expression regulation on cell cycle, apoptosis and invasiveness of cervical squamous cell carcinoma Hela229.
Method
1. the LRIG3 antisense oligonucleotide (LRIG3ASODN), the just oligonucleotide sequence (LRIG3SODN) and the missense oligonucleotide sequence (LRIG3MSODN) were designed and synthesized. The transfected Hela229 cells were transfected with liposome Lipofectamine2000 as the transfected cells. The cells transfected with liposomes were used as blank control cells.
The expression of 24h, 48h and 72h after transfection of 24h, 48h and 72h was detected by 2. Western blot (150 g/ml, 200, g/ml, 250 mu g/ml) LRIG3ASODN, LRIG3MSODN and LRIG3SODN.
3. RT-PCR was used to detect the expression of 24h and 48h and 72h after transfection at different concentrations (150 g/ml, 200 g/ml, 250 g/ml) LRIG3ASODN, LRIG3MSODN and LRIG3SODN.
4. the cell proliferation was detected by MTT, cell apoptosis was measured by flow cytometry, cell adhesion test was used to detect cell adhesion ability, and cell invasion ability was detected by Transwell chamber method.
Result
The expression of LRIG3 protein and mRNA in group 1. ASODN was significantly lower than that in group SODN, in group MSODN and in blank control group (P0.05), and there was a concentration dependence and time dependence. The expression of LRIG3 decreased with the increase of transfection concentration and the prolongation of time.
The expression of EGFR protein and mRNA in group 2. ASODN was significantly higher than that in group SODN, and in MSODN group, the blank control group was significantly increased (P0.05). The expression of.EGFR and LRIG3 expressed a negative correlation with the increase of the transfection concentration and the prolongation of the expression of EGFR.
3. MTT test showed that the proliferation rate of Hela229 cells in group LRIG3ASODN was significantly higher than that of other groups (P0.05). With the increase of LRIG3ASODN transfection concentration, the expression inhibition time was prolonged and the cell proliferation rate was significantly increased, the difference was statistically significant (P0.05).
4. the apoptosis rate of Hela229 cells decreased significantly after transfection of LRIG3ASODN. With the increase of transfection concentration and the prolongation of time, the apoptosis rate of cells decreased.
The adhesion test of 5. cell matrix showed that the adhesion rate of Hela229 cells increased obviously after transfection of LRIG3ASODN. With the increase of transfection concentration, the time was prolonged and the adhesion rate of cells increased.
6. Transwell test showed that the migration ability of Hela229 cells increased significantly after transfection of LRIG3ASODN. The number of membrane cells in ASODN group (246 + 10) was significantly higher than that in group SODN (168 + 10), MSODN group (177 + 8) and blank control group (176 + 6).
The third part is the effect of LRIG3 expression regulation on the growth of human cervical squamous cell carcinoma xenografts in nude mice.
Method
1 x 107 Hela229 cells were injected into nude mice to construct nude mice model of nude mice, and the growth state of nude mice was observed after subcutaneous injection of LRIG3ASODN into the tumor. The expression of LRIG3 and EGFR protein and mRNA in the transplanted tumor of nude mice was measured by.Western blot and RT-PCR.
Result
In 1. ASODN LRIG3 group, the tumor volume in nude mice was significantly higher than that in the other two groups on the 18-30 day after injection (P0.05).
2. Western blot and PT-PCR detected the expression of LRIG3 protein and mRNA in nude mice xenograft. The expression of LRIG3 in ASODN group was significantly lower than that in MSODN and blank control group (P0.05).
3. Western blot and PT-PCR detected the expression of EGFR protein and mRNA in nude mice xenograft. The expression of EGFR in ASODN group was significantly higher than that in MSODN and blank control group (P0.05).
conclusion
1. the expression of LRIG3 in cervical squamous cell carcinoma tissues and CIN tissues is low, and the expression of EGFR is highly expressed. The expression of LRIG3 is closely related to the invasion and metastasis of cervical squamous cell carcinoma. It suggests that the negative feedback regulation of EGFR plays a role in inhibiting the development of cervical squamous cell carcinoma.
2. LRIG3ASODN can significantly reduce the expression of LRIG3 protein and mRNA in the Hela229 cells of cervical squamous cell carcinoma, and increase the expression of EGFR and mRNA, which can inhibit the apoptosis of cervical squamous cell carcinoma cells and enhance the invasive ability of the cells, suggesting that LRIG3 plays an important role in the proliferation, apoptosis and invasion of cervical squamous cell carcinoma cells.
3. LRIG3ASODN could lead to accelerated tumor formation and increased volume of cervical squamous cell carcinoma in nude mice. The expression of LRIG3 protein and mRNA in the transplanted tumor tissue was significantly decreased, and the expression of EGFR protein and mRNA was increased.
【学位授予单位】：郑州大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R737.33
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