| BackgroundThe treatment of malignant tumors is still a challenge. Cancer cells proliferate quickly, resist the apoptosis, evade the immune system and metastasis easily. The mortality of liver cancer is in the third place of malignant tumors all over the world currently.The invasiveness and metastasis of tumors is a significant cause for the patients’ death. Research on hepatocellular carcinoma in the past mainly focus on the following areas:①molecular epidemiology:infections of hepatitis B virus,hepatitis C virus,etc;②high risk factors which caused hepatocellular carcinoma:alcohol abuse, and yellow Aspergillus toxins pollution;③Molecular genetics, such as the structure deformity of chromosome1th;④gene molecular biology:the abnormalities of N-ras, p53, p21,etc;⑤epigenetics:the expression of miR-221and miR-222, miRNA (microRNA,Micro-RNA) in tumor tissue is abnormal,which is related with invasion of HCC.⑥the abnormalities lof signaling pathway.Recent research found that Non-coding RNA found long non-coding RNA.IncRNA.may regulate protein coding gene expression in the levels of transcription and translation.One or more IncRNA with relevant regulation function could be found in most of the genes encoding.This provides an entirely new idea for disease research.However currently research on IncRNA in hepatocellular carcinoma is hardly to seen.Past studies found significantly elevated expression of IncRNAUC001kfo in hepatocellular carcinoma. Predictive analysis shows the target gene of UC001kfo may be ACTA2, the protein product of which is a-smooth muscle actin (a-smooth muscle action,a-SMA).a-SMA may be involved in the regulation of invasion and metastasis of liver cancer. Recently,the research about IncRNA is less in liver cancer. Previous experimental studies have found that IncRNAUC001kfo expression was significantly increased in liver cancer. Analysis of the target gene of UC001kfo may be ACTA2, its protein expres-sion is a-smooth muscle actin (a-smooth muscle the action, a-SMA), predictived that it may involve in the regulation of invasion and metastasis of liver cancer.A-SMA is a part of the cytoskeleton which participate in the division,movement, migration, morphology, maintenance, growth, and other important cellular activity of cells. a-SMA is to Promote the invasion and metastasis of liver cancer. Previous studies only use immunohistochemical methods to get a-SMA has different expression in the early hepatocellular carcinoma and later hepatocellular carcinoma with portal vein tumor thrombus. UCOOlkfo is newly discovered. The study using reverse transcriptase polymerase chain reaction (RT-PCR), in situ hybridization and immunohistochemical methods to discuss the realition of α-SMA、ACTA2、UC001kfo the and the metastasis invasion of liver cancer. It is proving whether a-SMA has different expression in the different periods of liver cancers, and prove whether UC001kfo and ACTA2expression is consistency,and the relation between long non-coding RNAUC001kfo and metastasis of liver cancer, to provide a new therapy of the treatment of liver cancerbasis.Materials and methods1.collection control liver tissue of15cases, cirrhotic li-ver tissue15cases, liver cancer tissue of30cases, adjacent tissues of25cases, including10cases of hepatocellular carcinoma wit-h portal vein tumor thrombus,15cases of control liver tissue from the liver emangioma, liver cyst surrounding normal liver organizations, All liver tissues were confirmed by pathology diagnosis and collect-ed from the department of pathology, the First Affiliated Hospital of Zhengzhou University from January2010to August2011, HCC tissues in accordance with the pathological differentiation are divided into three groups:low differentiation group, medium differentiation group and high differentiation group.All the tissue don’t limit the age and sex.2.Extracting the total RNA of liver cancer and reversing transcription them into cDNA, then by PCR Amplification become them into cDNA, each PCR internal reference control are all GAPDH, getting the electrophoresis photographs, optical density scanning using the Image J software, the relative expression of a-SMA mRNA in strength are the ratio of the absorbance values of a-SMA mRNA and GAPDH absorbance values, finally comparing the differences in expression intensity.3.For paraffin sections collected, to conducted ACTA2and UC001kfo in situ hybridization in accordance with the instruction’s step and immunohistochemi-stry of alpha-SMA, Using BioSens Digital Imaging the System to analysis th-e situ hybridization image gray value.4.Test methods:Statistical analysis using SPSS17.0,data using mean±standard deviation, two comparion to take the t-test,between many groups to take variance analysis.Result1.The results of RT-PCR show that a-SMA mRNA are positive expression in all HCC tissues.Control liver group and the liver cancer group of a-SMA mRNA expre-ssion were0.24±0.01,0.62±0.03, t test results show that the difference between the two groups was thought to be statistically significant (P<0.01).2. The results of immunohistochemistry show that α-SMA expression in co-ntrol liver, liver cancer tissue and portal vein tumor thrombus organization values are125.69±29.17,187.43±17.66,218.70±13.52respectively, variance analysis shows that the difference of a-SMA expression has statistical significance(P<0.01), the expression of portal vein tumor thrombus group is the highest; In accordance with the liver pathology, liver tissues are divided into three groups:low differentiation group, medium differentiation group and high differentiation group, the expression values are237.14±25.83,190.57±19.90,165.67±22.64respectively, variance analysis shows that the expression difference of α-SMA has statistical significance (P<0.01), the expression of low differentiation group is the highest.2The results of.in situ hybridization show that UCOOlkfo in the group of liver cirrhosis are no expression or low expression, in the other three groups are all positive expression,these four groups of expression values are113.30±11.79,137.59±6.23,148.78±8.23,160.28±9.47respectivly. Variance analysis show the difference of UCOOlkfo expression has statistical significance (P<0.01), Differences among the four groups has statistical significance (P<0.01), respectively,the cirrhosis group<adjacent tissues<Liver cancer<.portal vein tumor thrombus group. Expression for ACTA2are109.89±9.74,125.22±32.16,149.06±8.43,156.57±8.86respectively. the difference of ACTA2expression has statistical significance (P<0.01),difference among the four groups has statistical significance (P<0.05),the expression are cirrhosis group<.adjacent tissues.<Liver cancer <.portal vein tumor thrombus group respectively.Conclusion1.the expression of α-SMA in liver cancer increases significantly, especially portal vein tumor thrombus group and the low differentiated group, It is related to the invasion and metastasis of liver cancer.2. the expression of UC001kfo and ACTA2in hepatocellular carcinoma increase significantly, especially the portal vein tumor thrombus group, UC001kfo may regulate the expression of ACTA2to promte invasion and metastasis of liver cancer... |
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