Li Bao-ding, Zhao Qing-chuan, Zhu Yang-ting, Zhang Fu-qin, Dou Ke-feng
Department of Hepatobiliary Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
Zhonghua Wai Ke Za Zhi. 2006 Apr 1;44(7):458-62.
To investigate the expressions of glypican-3 (GPC3) mRNA in hepatocellular carcinoma (HCC) tissues and peripheral blood cells (PBCs), and to determine the values of GPC3 mRNA in the diagnosis of HCC and HCC micrometastasis.
Using semi-quantitative and nested reverse transcription polymerase chain reactions (RT-PCR), we detected the expressions of AFP and GPC3 genes in the tissues of 41 HCC, 41 paracancer and 52 non-HCC liver samples (41 far from HCC tissues and 11 normal liver tissues), and in the PBCs of 67 specimens from subjects.
The semi-quantitative RT-PCR displayed GPC3 mRNA was expressed in all samples of tissues and PBCs, and the relative intensities of its expressions in HCC, paracancer, non-HCC liver tissues were 78.9 +/- 35.5, 30.6 +/- 21.6, 23.8 +/- 15.5 respectively. The AFP mRNA expression values were 61.2 +/- 32.6, 31.5 +/- 23.6, and 21.2 +/- 15.9 respectively. The expression of each gene in HCC differed significantly from those in other two kinds of tissue samples (P < 0.01). The expressions of GPC3 mRNA and AFP mRNA, accounting for 80.5% and 63.4% in all the HCC tissues, were higher than their respective peak values in the tissues of non-HCC liver (+1.96s), but the expressions of at least one of the two genes was elevated in 92.7% of all the HCC tissues. There was a significant difference between combined detection of two genes and single AFP mRNA detection in HCC tissues (P < 0.01). Clinicopathologically, AFP mRNA was related with the grade of HCC and serum AFP, while GPC3 mRNA was related with not only the grade of HCC but also the invasion of HCC. The relative intensities of GPC3 mRNA expressions in PBCs of 67 specimens was 15.9 +/- 9.0, and GPC3 mRNA expressed in three kinds of tissue samples were all stronger than its counterparts in PBCs (P < 0.01). The GPC3 mRNA expression values in PBCs of the HCC group and the non-HCC group were respectively 16.1 +/- 8.3, 15.6 +/- 10.2, there was no significant difference between the two groups. Of the HCC metastasis group and the HCC non-metastasis group, the respective GPC3 mRNA expression values in PBCs were 16.0 +/- 9.0 and 16.3 +/- 7.7, there was also no significant difference between the two groups. The nested RT-PCR showed that the positive rates of AFP mRNA expressions in PBCs from the HCC group and the non-HCC group were 56.1% and 23.1%, and the difference between the two groups was significant (P = 0.011). The positive rates of AFP mRNA expressions in PBCs from the HCC metastasis group and the HCC non-metastasis group were 80.9% and 30.0%, and there was also a significant difference between the two groups (P = 0.002).
Although GPC3 mRNA is expressed broadly, it still may serve as a potential tissue biomarker in the diagnosis of HCC. Detecting the expression of the two genes in the tissues will improve the screening and diagnosis of HCC. GPC3 is prevalently transcribed in the PBCs, but we have not found any relationship between the GPC3 expression in PBCs and the metastasis or recurrence of hepatocellular carcinoma, thus we can not identify HCC micrometastasis with GPC3 mRNA.
研究磷脂酰肌醇蛋白聚糖-3(GPC3)mRNA在肝细胞癌(HCC)组织及外周血细胞(PBCs)中的表达情况,探讨GPC3 mRNA在HCC诊断及微小转移中的价值。
采用半定量及巢式逆转录聚合酶链反应(RT-PCR),检测41例HCC组织、41例癌旁组织及52例非HCC肝脏组织(41例远离HCC组织及11例正常肝脏组织)以及67例受试者PBCs中AFP和GPC3基因的表达。
半定量RT-PCR显示GPC3 mRNA在所有组织及PBCs样本中均有表达,其在HCC、癌旁、非HCC肝脏组织中的相对表达强度分别为78.9±35.5、30.6±21.6、23.8±15.5。AFP mRNA表达值分别为61.2±32.6、31.5±23.6、21.2±15.9。各基因在HCC中的表达与其他两种组织样本差异有统计学意义(P<0.01)。GPC3 mRNA和AFP mRNA在所有HCC组织中的表达分别占80.5%和63.4%,高于非HCC肝脏组织中的各自峰值(+1.96s),但在92.7%的HCC组织中至少有一个基因表达升高。HCC组织中两基因联合检测与单一AFP mRNA检测差异有统计学意义(P<0.01)。临床病理方面,AFP mRNA与HCC分级及血清AFP相关,而GPC3 mRNA不仅与HCC分级相关,还与HCC侵袭相关。67例标本PBCs中GPC3 mRNA相对表达强度为15.9±9.0,其在三种组织样本中的表达均强于PBCs中的表达(P<0.01)。HCC组与非HCC组PBCs中GPC3 mRNA表达值分别为16.1±8.3、15.6±10.2,两组间差异无统计学意义。HCC转移组与非转移组PBCs中GPC3 mRNA表达值分别为16.0±9.0和16.3±7.7,两组间差异亦无统计学意义。巢式RT-PCR显示,HCC组与非HCC组PBCs中AFP mRNA阳性率分别为56.1%和23.1%,两组间差异有统计学意义(P = 0.011)。HCC转移组与非转移组PBCs中AFP mRNA阳性率分别为80.9%和30.0%,两组间差异也有统计学意义(P = 0.002)。
尽管GPC3 mRNA广泛表达,但仍可能作为HCC诊断的潜在组织生物标志物。检测组织中两基因表达可提高HCC的筛查及诊断。GPC3在PBCs中普遍转录,但未发现PBCs中GPC3表达与肝细胞癌转移或复发的关系,因此不能用GPC3 mRNA鉴定HCC微小转移。