[Comparison of quantitative detection of BCR::ABL1 p210 transcript levels: a multicenter study].

Objective: To assess the capability of seven reference medical laboratories to detect BCR::ABL1 p210 transcription levels and to compare the results among those laboratories. Methods: The interlaboratory comparison was carried out in two stages. The samples were prepared by the reference laboratory. The quantitative values of BCR::ABL1 p210 of the comparison samples covered 0.001%-0.01%, 0.01%-0.1%, 0.1%-1%, 1%-10% and>10% in each stage. Real-time quantitative PCR (RT-PCR) and dPCR (digital PCR) were used to examine the samples. The conversion factor (CF) was calculated and validated for each laboratory. Results: In the RT-PCR comparison, one laboratory was failed to detect BCR::ABL1 p210 in fourteen samples at the first stage. The results of the other six laboratories were qualified with the bias <±1.2 folds (-0.133-0.338) and 95% limits of agreement within ±5 folds (upper limit 0.147-0.785, lower limit -0.770--0.109), and the corresponding CF values were calculated and validated. In the dPCR comparison, one laboratory did not report results at the second stage. The results of the other six laboratories were qualified with the bias <±1.2 folds (-0.026-0.267) and 95% limits of agreement within±5 folds (upper limit 0.084-0.991, lower limit -0.669--0.135), and the corresponding CF values were calculated and validated. The samples with BCR::ABL1 p210 quantitative values of 0.01%-0.1%, 0.1%-1%, 1%-10% and >10% could be detected by both RT-PCR and qPCR. When the quantitative value of BCR::ABL1 p210 was 0.001%-0.01%, the detection rate of dPCR was higher than that of RT-PCR (85.56% vs. 68.00%). Conclusions: A good consistency is present among various laboratories. The quantitative value of BCR::ABL1 p210 is comparable among laboratories as shown by the CF value conversion. For quantitative detection of BCR::ABL1 p210 deep molecular reaction, dPCR has a higher positive detection rate and more advantages than RT-PCR. To ensure the accuracy and reproducibility of the BCR::ABL1 p210 test, it is imperative for every laboratory to enhance their daily quality control practices.

Variation of ER status between primary and metastatic breast cancer and relationship to p53 expression*.

OBJECTIVE: Estrogen-dependent growth of breast cancer can be blocked by anti-estrogens. Estrogen receptor (ER) presence in breast cancer implies responsiveness to endocrine therapy. However, for those patients who ultimately develop resistance to endocrine therapy, the mechanisms remain unclear. The present study attempted to compare the expression status of ER mRNA in a series of primary breast tumors with matched metastases and explored the relation between ER and mutant p53 expression. METHODS: In situ hybridization using a digoxigenin-labeled estrogen receptor cDNA probe was employed to determine the expression of ER mRNA in 52 cases of primary tumors and their matched axillary lymph node metastases. Immunohistochemical staining using a monoclonal antibody against ER was also performed. RESULTS: ER expression was observed in 53.8% (28/52) of primary tumors and 48% (25/52) of metastases, while 57.7% (30/52) of primary tumors and 53.8% (28/52) of metastases showed ER mRNA positivity. There were variations in ER status between in situ hybridization and immunohistochemistry measurements and between primary tumors and metastases. Mutant p53 expression was inversely associated with ER-negative, high-grade tumors. CONCLUSIONS: In situ hybridization may be a more specific and sensitive method for determination of ER status than immunohistochemistry. It is possible that the biologic properties of ER change, and these changes may influence tumor response to endocrine therapy. In view of the ER variation, it was suggested that the ER status of metastatic tumors in addition to primary tumors should be taken into consideration in order to better determine the benefit of clinical endocrine therapy.

Clinicopathological risk factors and prognostic evaluation in hepatocellular carcinoma recurrence after surgery.

AIM: To analyze the clinicopathological risk factors in hepatocellular carcinoma recurrence after surgery. METHODS: We used significance testing (χ(2) and Student's t-test) of single and multiple factors, and Wilcoxon Cox tropic examination; a retrospective clinicopathological analysis was performed on 156 cases of hepatocellular carcinoma after hepatectomy. RESULTS: Of the 156 cases, 68.4%, 57.3%, 46.7%, 31.5%, and 28.6% had one, two, three, four, and five postoperative tumor-free years, respectively; the total recurrence rate was 53.2% (83/156). In the 83 recurrent cases, 65 were intrahepatic subclinical, with a resection rate of 78.3% (65/83). The relevant factors involved in recurrence were: male gender, tumor number and size, capsule infiltration, and portal vein involvement. These factors were an obvious influence on the prognosis of the patients with postoperative hepatocellular carcinoma (P < 0.05). In the recurrent liver carcinomas, 63.1% of tumor nodes (41/65) were at the ipsilateral segment of the primary tumor nodes. CONCLUSION: Male gender, tumor number and size, capsule infiltration, and portal vein involvement are factors for postoperative hepatocellular carcinoma recurrence. Recurrence is mainly unicentral. The right front liver lobe is the segment with a high rate of recurrence.

[Expression and significance of oncoprotein p53, rasp21 and p185 in carcinomas and polyps of the large intestine].

Seventy eight cases of carcinoma and 14 cases of poly of large intestine were examined to observe the expression of p53, rasp21 and p185 oncoproteins with immunogold silver staining method. The positive expressions for p53, rasp21 and p185 were 53.9%, 46.2% and 51.3% respectively. But only poorly positive expression was found in large intestine polyps for p185 staining. The difference between the carcinomas and polyps for p185 staining was significant (P < 0.01). The normal mucoepithelium near the cancer region expressed positive staining for p53, rasp21 and p185 were 0%, 2.6% and 5.1% respectively. These results were also significantly different when compared to cancer tissue (P < 0.01). In addition, of these 78 cases, 33 cases (42.3%) gave coordinately positive expression for p53 and rasp21 oncoprotein; p53 positive expression was obviously stronger in poorly differentiated carcinoma and mucoadenocarcinoma than in highly and moderately differentiated adenocarcinoma (P < 0.01).

[Interstitial collagen distribution and its mRNA expression in human primary liver cancer].

The localization of type I collagen and type III procollagen in 29 cases of hepatocellular carcinoma (HCC) and 2 cases of cholangiocarcinoma as well as their surrounding tissues were studied with ABC method. The results showed that the distribution of these two types of collagen was similar in PLC as well as in its surrounding tissues. The distribution of interstitial collagen in HCC might be classified into two patterns by the authors' namely, the vasiform and non-vascular distributions. The carcinoma with vasiform distribution pattern tends to grow infiltratively with much higher rate of portal venous embolism and intrahepatic metastasis development than that of the non-vascular pattern (P < 0.05). Employing the in situ hybridization technique for type I collagen mRNA, the transcription of collagen mRNA occurred not only in the interstitial cells but also in some of the liver carcinoma cells and giant tumor cells. No positive signals of type I collagen mRNA were noticed in the non-tumorous surrounding tissues.

A 54-kDa protein overexpressed by chloroquine-resistant Plasmodium berghei ANKA strain.

Using an insoluble chloroquine-adsorbent, a 54-kDa protein (with a range of 50-60 kDa) was extracted from serum of mice infected with chloroquine-resistant (CR) Plasmodium berghei ANKA strain. Immunoblotting assay with antiserum against the 54-kDa protein showed that the content of the protein was higher in serum of mice infected with the CR parasites than that of mice infected with chloroquine-sensitive (CS) P berghei ANKA strain, and that instead of the 54-kDa protein, a set of 15-, 16-, and 23-kDa proteins was found to be highly overexpressed in lysate of purified CR parasites in comparison with that of purified CS parasites, suggesting the 54-kDa protein probably to be composed of 3 subunits. These findings may bear great importance in probing mechanism of chloroquine resistance in malaria parasites.