| Background:Gastric cancer is one of the most common malignant tumors, whose incidence in the world ranked fourth for the malignant tumor, the mortality rate in the second place.In our country, about 300000 new cases occur every year, constituting a serious threat to the health.And metastasis are the most common cause of death in patients with gastric cancer, the molecular mechanism of it has been a hotspot in the field of cancer research.Micro RNA(mi RNA) is a large family of endogenous, highly conserved during evolution of single small non-coding RNA, involved in the regulation of human body various life activities such as cell proliferation, differentiation, apoptosis, etc. The expression of mi RNA disorders can lead to many diseases, and have a very close relationship with the occurrence and development of a variety of malignant tumor. In recent years, many studies have shown that the phenomenon of mi RNA’s disorder exists in gastric cancer, illustrating mi RNA involved in the occurrence of gastric cancer.Our study will preliminarily research and explore the expression level of mi R-29a-3p in gastric cancer cell line, biological functions and possible mechanism on the basis of preliminary study. Objective:To preliminarily explore how hsa-mi R-29a-3p( mi R-29a) affects gastric cancer peritoneum metastasis and its mechanism research. Methods:1.q RT-PCR was used to verify the expression level of mi R-29a-3p in three groups of gastric cancer cell line and high metastasis cell line: GC9811/GC9811-P, MKN28-M/MKN-28 NM, SGC7901-M/SGC7901-NM.2.The lentivirus vector of over-expressed mi R-29 a and its relative negative control were transferred in GC9811-P; On the contrary, the lentivirus vector inhibiting the expression of mi R-29 a and its relative negative control were transferred in GC9811.3.The MTT assay, cloning-forming assay, Transwell, wound healing assays and flow cytometer were then performed to investigate cell proliferation, migration and apoptosis capabilities.4.Bioinformatic method was used to predict out the potential target, and q RT-PCR and western blot were used to verify it.5.si RNA and empty vector were transfected to silence the expression of HAS3 to future verify its functions and verify whether it’s the target of mi R-29a-3p. Result:1.the expression of mi R-29 a was higher in high metastatic cell lines(GC9811-P、MKN28-M、SGC7901-M) than it in low metastatic cell lines.2.Successfully construct cell lines of high and low expression of mi R-29a-3p,and q RT-PCR verified the success of transfection.3.Over-expressing mi R-29 a could suppress the ability of cell proliferation and migration but promote apoptosis; On the contrary, inhibiting mi R-29 a could promote the ability of cell proliferation and migration while suppress apoptosis.4.Bioinformatics predictions suggest HAS3 may be one of possible target genes of mi R-29a-3p, q RT- PCR results showed that in the up-regulating mi R-29 a cell line, the expression of HAS3 has a tendency to reduce but without statistical significance; in the down-regulating mi R-29 a cell line, the expression of HAS3 has a tendency to increase but without statistical significance. According to the results of Western blot, the expression of HAS3 decreased when up-regulating mi R-29 a while the expression of HAS3 increased when down-regulating mi R-29 a.5.Knockdown of HAS3 inhibits the proliferation and migration of gestric cancer,illustrating HAS3 promote the proliferation and metastasis of gastric cancer cells. Knockdown of HAS3 inhibits the proliferation and migration of mi R-29 a high-expressed cell line,which has the similar result of over-expressing mi R-29 a. The above two instructs mi R-29 a can negatively regulate HAS3, and confirms HAS3 may be one of the potential target genes of mi R-29a-3p. Conclusion:mi R-29a-3p inhibits the proliferation and metastasis of gastric cancer cells and promoting apoptosis, and may play its role through one of its downstream target molecules HAS3. |
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