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Investigation of the Role of Heme Oxygenase-1 on Peritoneal Metastasis of Gastric Cancer and Its Involved Mechanisms

Author: HuiLiangLiang
Tutor: BaiFeiHu；FanDaiMing
School: Ningxia Medical University
Course: Internal Medicine
Keywords: Heme oxygenase -1 Peritoneal metastasis Cell proliferation Apoptosis Angiogenesis
CLC: R735.2
Type: Master's thesis
Year: 2010
Downloads: 41
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Abstract

Background heme oxygenase -1 (Heme oxygenase-1), also known as heat shock protein 32 (Heat-shock protein32), relative molecular weight of 32 kDa. Gene Name for HO-1, is located on human autosomal 22q13. HO-1 is the rate-limiting enzyme degradation of heme, control ferrous (Fe ^{2 ) metabolism. HO-1 is its role in cardiovascular disease, manifested as HO-1 to maintain cell homeostasis, reduce tissue oxidative damage, weaken the inflammatory response plays an important role in inhibition of apoptosis [1] . Recent studies have found that HO-1 and tumor cell proliferation, angiogenesis, apoptosis and biological behavior is closely related to the increased expression of HO-1 can promote the proliferation of tumor metastasis, by the turn of HO-1 SiRNA (HO-1 small interference RNA) 1 protein and tumor development are inseparable. Existing studies have shown that over-expression of HO-1 protein can promote human gastric cancer cells, tumor formation and proliferation of cancer cells, and the incidence of gastric cancer apoptotic factors P21 start [3]. About HO-1 protein in the peritoneal metastasis of human gastric cancer and its mechanism is not yet clear. the retroperitoneal metastasis rate (100%), can simulate the free cancer cells shed into the abdominal cavity after implantation, adhesion, invasion and growth around the tumor pathological angiogenesis step. GC9811-P cell lines compared to their maternal gastric cancer cell lines GC9811, the expression of HO-1 protein was significantly enhanced initially speculated that HO-1 protein in the peritoneal metastasis process plays an important role in further by Western blot Western blot . [Objective] To clear the HO-1 protein expression in human peritoneal metastasis of gastric cancer tissues and compare HO-1 protein in cancer tissue and normal tissue expression differences, explore the HO-1 protein in human gastric cancer peritoneal metastasis in patients with pathological parameters between relationship. 2, application of the HO-1 protein inhibitor peritoneal highly metastatic cell line HO-1 protein expression by comparing high and low expression of HO-1 protein cell proliferation, apoptosis, angiogenesis and biological behavior changes to clarify the HO-1 protein in the peritoneal metastasis process and its possible mechanisms. 【Methods】 1, immunohistochemistry SP method detected 68 cases of peritoneal metastasis of gastric adenocarcinoma tissue cancer and adjacent peritoneal tissue HO-1 protein expression, as well as 46 cases of peritoneal metastasis of gastric adenocarcinoma HO-1 protein expression, Western blotting assay peritoneal tissue next to the 14 cases of peritoneal metastasis of gastric adenocarcinoma of postoperative tissue with cancer HO-1 protein expression, as well as cell lines GC9811-P and GC9811, GC7901 cell line HO-1 protein expression. 2, MTT assay and 1 × 10 ^{-4 1 × 10 ^{-5 , 1 × 10 ^{to -6 1 × 10 ^{-7 , 1 × 10 ^{-8 , 1 × 10 ^{-9 mol/LHO-1 inhibitors were incubated 48 h peritoneal high transfer peritoneal highly metastatic cell HO-cell proliferation; flow cytometry low expression of HO-1 protein in peritoneal highly metastatic cell cycle and apoptotic index; Western blot Western blot method for the determination and inhibitor were incubated 1, Bcl-2, Bax protein expression levels. 3, chicken embryo allantoic membrane experimental observation were incubated with the inhibitor, high peritoneal metastatic cells capsular surface of the number of new blood vessels in the chick embryo number of blood vessel branch points, evaluation angiogenesis inhibition of HO-1 protein expression changes. [Results] 1, immunohistochemical staining results showed that HO-1 protein in gastric cancer peritoneal tissue positive expression rate was 41.2%, the positive expression rate in the the peritoneal tissue adjacent to metastatic carcinoma 0, and peritoneal metastasis gastric adenocarcinoma, the positive expression rate was 21.7%; HO-1 protein in human peritoneal metastasis tissue red positive expression rate was significantly higher than in adjacent peritoneal tissue (P lt; 0.01) and no peritoneal metastasis of gastric adenocarcinoma (P lt ; 0.05), the expression of HO-1 protein in the peritoneal metastasis of gastric cancer tissues was significantly enhanced. 2, immunohistochemical staining of HO-1 protein in the peritoneal metastasis tissue expression of strength and the patient's age, gender, lymph node metastasis, and metastatic carcinoma tissue differentiation: poorly differentiated metastatic tissues HO-1 protein expression strength significantly higher than moderately differentiated metastatic carcinoma tissue (P lt; 0.05), moderately differentiated metastatic carcinoma of HO-1 protein expression was significantly higher than the well-differentiated metastatic tissues (P lt; 0.05). Western blot blotting showed that HO-1 protein expression in human peritoneal metastasis of gastric cancer tissue specimens after level significantly higher than its paraneoplastic peritoneal tissue expression (P lt; 0.05), HO-1 protein in highly metastatic gastric cancer peritoneal cells was significantly higher than GC9811 cells and the GC7901 cell expression, and further validate the high expression of HO-1 protein in metastatic cancer tissues and cell lines. HO-1 protein inhibitors and peritoneal highly metastatic cells were incubated cell proliferation was measured by MTT method, highly metastatic peritoneal cell HO-1 protein expression by inhibiting proliferation reduced when the inhibitor concentration to 1 × 10 ^{-4 mol / L when the cell proliferation inhibition rate (62.04 ± 6.13)%, the the IC ₅₀ 1.2 × 10 ^{-5 mol / L, may play an important role in the proliferation of HO-1 protein in peritoneal highly metastatic cells. Inhibitor of HO-1 protein in gastric cancer peritoneal highly metastatic cells were incubated flow cytometry peritoneal highly metastatic cell apoptotic index and cell cycle, low expression of HO-1 protein highly metastatic peritoneal cells apoptotic index increased 48 h apoptotic index (61.1 ± 5.8)%, and cell cycle arrest in G1 and S phase, the period of 48 h cells (G1, S) (88.23 ± 6.7)%; Western immunoblot assay showed low expression of HO-1 protein high peritoneal metastatic cells accompanied by Bcl-2 and Bax protein expression levels decreased HO-1 protein is involved in peritoneal highly metastatic cell apoptosis, and the apoptosis and cell cycle G1, S arrest. 6, chorioallantoic membrane assay (CAMs): HO-1 inhibitors containing 2 × 10 ^{6 GC9811-P cells cultured in serum chorioallantoic membrane surface, cultured for 4-5d after the chick neovascularization and vascular branching points number was reduced (P lt; 0.05), low expression of HO-1 protein in gastric cancer peritoneal highly metastatic cell angiogenesis was significantly weakened, HO-1 protein in highly metastatic cells and peritoneal angiogenesis The closely related. [Conclusion] HO-1 protein highly expressed in the organization and its cell lines of human gastric cancer peritoneal metastasis, peritoneal metastasis tissue differentiation is lower the higher the intensity of its HO-1 protein expression; low expression of HO-1 protein in gastric cancer peritoneal highly metastatic cells proliferation, reduced the role of angiogenesis, increased apoptosis and cell cycle arrest (G1 / S phase), HO-1 protein may play an important role may be in the process of peritoneal metastasis.}}}}}}}}}}

Investigation of the Role of Heme Oxygenase-1 on Peritoneal Metastasis of Gastric Cancer and Its Involved Mechanisms

Abstract

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