| Objective:Exosome is produced within the cell, a size of about 30-140 nm membranous vesicles containing therein various types of nucleic acids, proteins, lipids. miRNA-221 and non-coding RNA Exosome embedded base pairing can promote epithelial-mesenchymal of Epithelial-to-mesenchymal transition(EMT) of tumor metastasis through more mediated signaling pathways. Therefore, detection of gastric cancer patients serum exosome content within miRNA whether as biological indicators to determine whether metastasis of gastric cancer staging and to guide treatment.Methods: We were collected from June 2014 to March 2016 Check ovarian metastasis of gastric cancer patients Hunan Provincial Tumor Hospital serum 30 parts(experimental group). Inclusion criteria are:(1) has been through the gastroscope, biopsy, EUS, CT, B super diagnosed with ovarian metastasis of gastric cancer patients;(2) found no other primary tumor;(3) patients not receiving chemotherapy Chinese medicine treatment and other auxiliary treatment;(4) the non-recurrence of gastric cancer patients. 30 parts of female patients without ovarian metastasis of gastric cancer serum(control group). Inclusion criteria are:(1) has been through the gastroscope biopsy, EUS, CT, B super diagnosed with stomach cancer;(2) not found gastric cancer metastasis;(3) found no other primary tumor;( 4) patients not receiving chemotherapy, Chinese medicine treatment and other auxiliary treatment;(5) the non-recurrence of gastric cancer patients. Serum samples drawn at 3000 g centrifuged for 15 min to remove cells or cell debris; the supernatant was transferred to another clean sterile tube, add the right amount of Exo Quick TC reagent; centrifuge tube upside down, mix; 4 ℃incubated overnight( at least 12 hours); room temperature or 4 ℃1500g centrifugal 30 min, bottom of the tube visible precipitate; removing the supernatant; 1500 g continue centrifugal 5min, carefully remove the upper layer of the liquid component; 100-500 ul buffer with the resuspended pellet. By western blot detection of CD9, CD63 confirmed exosome successfully extracted. TRIzol method of exsome in five kinds of miRNA(miR-221, miR-376 c, miR-744, miR-451, miR-486) were extracted. Then to compare experimental and control groups miRNA content is determined by PCR experiments.Results: In order to explore the gastric metastatic ovarian expression and non-metastatic patients with serum exosome of miRNA whether gastric cancer with ovarian metastasis were correlated, we selected five cancer patients serum exosome high expression of miRNA detection, found that compared in gastric cancer with ovarian metastasis in patients gastric cancer with ovarian metastasis in patients miRNA other exosome in embedded in does not occur in serum exosome of miR-221 content was significant difference(P <0.01), gastric cancer ovarian metastasis in patients with serum exosome of miR-221 content was significantly higher expression, such as miR-376 c, miR-774, miR-451 and miR-486 in patients with metastatic ovarian and gastric cancer patients with non-metastatic serum levels no significant difference. For a more detailed analysis to serum exosome of miR-221 in the age group of ovarian metastasis of gastric cancer patients with different differences, ovarian metastasis in patients with gastric cancer will be divided by age 20-40 years, 40-60 years, 60 years more than three groups, of which 20-40-year-old patients serum exosome compared to other age groups, and miR-221 showed higher expression levels. These findings suggest that miR-221 in serum exosome content has great potential as a biomarker to diagnose gastric cancer with ovarian metastasis, especially for 20-40 year-olds ovarian metastasis of gastric cancer patients.Conclusions: 1. ovarian metastasis of gastric cancer patients with serum exosome in miR-221 expression was significantly higher. 2. The serum exosome of miR-221 as a diagnostic indicator of gastric cancer metastatic ovarian biology is very promising. 3. Ovarian metastasis of gastric cancer in patients with serum exosome of miR-221 content was also associated with age, the higher index in patients 20-40 years of age, suggesting a more significant diagnostic features. |
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