ABSTRACT
Objective@#To investigate the expression of HOXA terminal transcript antisense RNA (HOTTIP) in hepatocellular carcinoma (HCC) tissues and to explore its effect on proliferation, invasion and migration in HepG2 cells.@*Methods@#A total of 60 cases with HCC tissues undergoing excision surgery and 60 cases of corresponding paracancerous tissues from January 2012 to June 2018 in Dandong First Hospital of Liaoning Province were collected. The expressions of HOTTIP in HCC tissues and paracancerous tissues were detected by using real-time quantitative polymerase chain reaction (RT-qPCR), and the relationship between the expression and clinicopathological features was analyzed. HepG2 cell line with high expression of HOTTIP constructed by cell transfer technique was treated as the experimental group, and the empty plasmid pcDNA3.1-NC was treated as the control group. The effect of HOTTIP on the proliferation of HepG2 cells was detected by using CCK-8 method, and the effect of HOTTIP on invasion and migration of HepG2 cells was detected by using Transwell assay.@*Results@#The expression of HOTTIP mRNA in HCC tissues was higher than that in paracancerous tissues, and there was no statistically significant difference (1.9±0.6 vs. 0.9±0.7, t = 6.069, P < 0.01). The whole HCC cases were divided into the high expression group (30 cases) and the low expression group (30 cases) according to the median value (1.92) of the expression of HOTTIP mRNA. The expression of HOTTIP was related with TNM stage, differentiation degree and lymph node metastasis (χ2 values were 10.800, 8.076, 5.711, all P < 0.05), but not with age, gender, tumor diameter, number of tumors, hepatitis and alpha fetoprotein (AFP) levels (all P > 0.05). The results of RT-qPCR showed that the expression of HOTTIP mRNA in HepG2 cells was increased after transfection of overexpressed HOTTIP and the differences was statistically significant compared with the control group (63±6 vs. 13±9, t = 9.129, P < 0.01). The results of CCK-8 method showed that the proliferation activity of cells was enhanced after the overexpression of HOTTIP in HepG2 cells (24 h, 36 h, 72 h at 490 nm absorbance was 1.497 ± 0.017 vs. 0.826±0.006, 2.002±0.025 vs. 1.211±0.020, 3.257±0.042 vs. 1.772±0.021), and the differences were statistically significant (t values were 5.321, 7.349, 8.793, all P < 0.01). In Transwell invasion assay, the number of cells transferred into the basement membrane in the experimental group was more than that in the control group (101±9 vs. 41±11), and the difference was statistically significant (t = 6.839, P < 0.01). In Transwell migration assay, the number of cells transferred into the basement membrane in the experimental group was more than that in the control group (112±9 vs. 53±11), and the difference was statistically significant (t = 7.105, P < 0.01).@*Conclusion@#The expression of HOTTIP in HCC tissues is up-regulated, and the overexpression of HOTTIP can promote the proliferation, invasion and migration of HCC cells.
ABSTRACT
Objective To investigate the expression of HOXA terminal transcript antisense RNA (HOTTIP) in hepatocellular carcinoma (HCC) tissues and to explore its effect on proliferation, invasion and migration in HepG2 cells. Methods A total of 60 cases with HCC tissues undergoing excision surgery and 60 cases of corresponding paracancerous tissues from January 2012 to June 2018 in Dandong First Hospital of Liaoning Province were collected. The expressions of HOTTIP in HCC tissues and paracancerous tissues were detected by using real-time quantitative polymerase chain reaction (RT-qPCR), and the relationship between the expression and clinicopathological features was analyzed. HepG2 cell line with high expression of HOTTIP constructed by cell transfer technique was treated as the experimental group, and the empty plasmid pcDNA3.1-NC was treated as the control group. The effect of HOTTIP on the proliferation of HepG2 cells was detected by using CCK-8 method, and the effect of HOTTIP on invasion and migration of HepG2 cells was detected by using Transwell assay. Results The expression of HOTTIP mRNA in HCC tissues was higher than that in paracancerous tissues, and there was no statistically significant difference (1.9±0.6 vs. 0.9±0.7, t=6.069, P<0.01). The whole HCC cases were divided into the high expression group (30 cases) and the low expression group (30 cases) according to the median value (1.92) of the expression of HOTTIP mRNA. The expression of HOTTIP was related with TNM stage, differentiation degree and lymph node metastasis (χ2 values were 10.800, 8.076, 5.711, all P<0.05), but not with age, gender, tumor diameter, number of tumors, hepatitis and alpha fetoprotein (AFP) levels (all P>0.05). The results of RT-qPCR showed that the expression of HOTTIP mRNA in HepG2 cells was increased after transfection of overexpressed HOTTIP and the differences was statistically significant compared with the control group (63±6 vs. 13±9, t=9.129, P<0.01). The results of CCK-8 method showed that the proliferation activity of cells was enhanced after the overexpression of HOTTIP in HepG2 cells (24 h, 36 h, 72 h at 490 nm absorbance was 1.497 ± 0.017 vs. 0.826 ±0.006, 2.002 ±0.025 vs. 1.211 ±0.020, 3.257 ±0.042 vs. 1.772 ±0.021), and the differences were statistically significant (t values were 5.321, 7.349, 8.793, all P < 0.01). In Transwell invasion assay, the number of cells transferred into the basement membrane in the experimental group was more than that in the control group (101 ±9 vs. 41 ±11), and the difference was statistically significant (t= 6.839, P< 0.01). In Transwell migration assay, the number of cells transferred into the basement membrane in the experimental group was more than that in the control group (112 ±9 vs. 53 ±11), and the difference was statistically significant (t=7.105, P<0.01). Conclusion The expression of HOTTIP in HCC tissues is up-regulated, and the overexpression of HOTTIP can promote the proliferation, invasion and migration of HCC cells.
ABSTRACT
Objective To investigate the expressions of Wnt3a and Wnt5a in papillary thyroid carcinoma (PTC) and their clinical significances. Methods Immunohistochemical SABC method was used to detect the expressions of Wnt3a and Wnt5a proteins in PTC tissues and their paracancerous tissues collected from 79 patients in Dandong First Hospital from January 2014 to June 2018, and the relationships between the expressions of Wnt3a and Wnt5a proteins and clinicopathological features of PTC patients were analyzed. The expressions of Wnt3a and Wnt5a proteins in 10 pairs of fresh PTC tissues and paracancerous tissues were detected by Western blot. Results The results of immunohistochemistry showed that the positive expression rates of Wnt3a and Wnt5a proteins in PTC tissues were significantly higher than those in paracancerous tissues [69.6％ (55/79) vs. 25.3％ (20/79), 60.8％ (48/79) vs. 20.2％ (16/79)], and the differences were statistically significant (χ 2 values were 31.092 and 26.894, both P < 0.01). The results of Western blot showed that the expressions of Wnt3a and Wnt5a proteins in 10 pairs of fresh PTC tissues was significantly higher than those in paracancerous tissues(1.61±0.40 vs. 0.43±0.14, 1.38±0.291 vs. 0.36±0.13), and the differences were statistically significant (t values were 16.234 and 13.493, both P < 0.01). The expressions of Wnt3a and Wnt5a in PTC tissues were correlated with TNM stage, differentiation, extramembranous invasion and lymph node metastasis (Wnt3a: χ2 values were 6.645, 15.945, 8.783 and 11.220; Wnt5a: χ2 values were 21.525, 7.611, 17.880 and 12.581, all P < 0.05), but not with patients'age, sex and tumor diameter (all P > 0.05). There was a positive correlation between Wnt3a and Wnt5a proteins expressions in PTC (r = 0.597, P < 0.01).Conclusion The abnormal expressions of Wnt3a and Wnt5a proteins in PTC may be related to the development of PTC.